user manual
Hoefer SE250
Mini-vertical gel electrophoresis unit
um SE250-IM/Rev.B0/04-12
Contents
Important Information ............................................ii
Waste Electrical and
Electronic Equipment (WEEE) ...............................vii
1. Gel Electrophoresis Unit
Function and Description ...................................1
2. Specifications ...................................................2
3. Operating Instructions ........................................4
4. Care and Maintenance .....................................14
5. Troubleshooting ...............................................15
Appendix ............................................................ 17
Bibliography ........................................................19
Ordering Information ............................................20
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Important Information – English
• If this equipment is used in a manner not specied
by Hoefer, Inc. the protection provided by the
equipment may be impaired.
• This instrument is designed for indoor laboratory
use only.
• Only accessories and parts approved or supplied
by Hoefer, Inc. may be used for operating,
maintaining, and servicing this product.
• Only use a power supply that is CE marked or
safety certied by a nationally recognized testing
laboratory.
• The safety lid must be in place before connecting
the power supply leads to a power supply.
• Turn all power supply controls o and disconnect
the power leads before removing the safety lid.
• Circulate only water or 50/50 water/ethylene glycol
through the heat exchanger if so equipped. Do
not connect the heat exchanger to a water tap or
any coolant source where the water pressure is
unregulated.
• Never introduce antifreeze or any organic solvent
into any part of the instrument. Organic solvents
will cause irreparable damage to the unit!
• Do not operate with buer temperatures above
the maximum specied technical specications.
Overheating will cause irreparable damage to the
unit!
Duležité Informace – Czech
• Pokud by toto zařízení je použito způsobem, který
není podle Hoefer, Inc. ochrana poskytovaná na
základě zařízení může být narušena.
• Tento nástroj je určen pro vnitřní použití v
laboratoři pouze.
• Pouze příslušenství a části schválen, nebo
poskytnutých Hoefer, Inc. mohou být použity pro
provoz, údržbu, a údržbě tohoto výrobku.
• zdroj napájení používají jen že je opatřen
označením CE osvědčena nebo bezpečnost
vnitrostátně uznanými zkušebními laboratoř.
• Bezpečnosti lid musí být zavedena před připojením
napájecí zdroj napájení vede k.
• Turn veškeré napájení kontroly vypnuto a odpojit
před odběrem energie vede bezpečnostní víko.
• Rozeslat pouze voda nebo 50/50 voda/
ethylenglykolu prostřednictvím výměník tepla je
li to vybavena. Nemají připojení výměník tepla s
vodními setřepná nebo jakékoli chladicí kapaliny
zdroje, kde tlak vody je neregulo.
• Nikdy zavést prostředek proti zamrznutí nebo
jakákoli organická rozpouštědla do jakékoli
části z tohoto nástroje. Rozpustidlům způsobí
nenapravitelné poškození jednotka!
• Nejsou provozována s pufru teplotách nad
maximální stanovenou technickými specikacemi.
Přehřátí způsobí nenapravitelné poškození
jednotka!
Vigtig Information – Danish
• Hvis dette udstyr bruges i en måde ikke
speciceret ved Hoefer, Inc. den beskyttelse, som
er blevet forsynet af udstyret kan måske svækkes.
• Dette instrument er designet for indendørs
laboratoriumbrug bare.
• Bare tilbehør og del godkendede eller forsynede
ved Hoefer, Inc. kan måske bruges for drive,
funktionsfejl, og betjening dette produkt.
• bruger Bare en strømforsyning, der er CE
markerede eller sikkerhed, som er blevet attesteret
af en, som nationalt er blevet anerkendt prøve
laboratorium.
• Sikkerhedlåget må være på plads før forbinding
strømforsyningsblyet til en strømforsyning.
• Drejer alle strømforsyningskontroller af og afbryder
kraftblyet før erning sikkerhedlåget.
• Cirkulerer bare vand eller 50/50 vand/ethylene
glykol gennem varmeveksleren i så fald udrustet.
Forbind ikke varmeveksleren til en vandhane
eller nogen kølemiddelkilde hvor vandtrykket er
unregulated.
• Introducerer Aldrig antifreeze eller noget organisk
opløsningsmiddel ind i nogen del af instrumentet.
Organiske opløsningsmidler vil forårsage uboelig
skade til enheden!
• Driver ikke med stødpudetemperaturer
over maksimummet specicerede tekniske
specications. Overheding vil forårsage uboelig
skade til enheden!
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Belangrijke Informatie – Dutch
• Indien deze uitrusting in een manier wordt
gebruikt die niet door Hoefer, Inc. is gespeciceerd
de bescherming die door de uitrusting is verzorgd
kan worden geschaad.
• Dit instrument is voor binnenlaboratoriumgebruik
enkel ontworpen.
• Enkel onderdelen en delen keurden goed of
leverden door Hoefer, Inc. kan voor het bedienen
worden gebruikt, handhavend en onderhouden
van dit product.
• gebruik Enkel een netvoeding die CE is markeerde
of veiligheid die door een is gecerticeerd die
nationaal is herkend testene laboratorium.
• Het veiligheidsdeksel moet in plaats voor het
verbinden van de netvoeding leidt tot een
netvoeding zijn.
• Doe alle netvoedingscontroles Uit en koppel los
de machtleiding voor het verwijderen van het
veiligheidsdeksel.
• Circuleer enkel water of 50/50 water/
ethyleenglycol door de hitte exchanger zo ja
uitrust. Verbind de hitte exchanger naar een
waterkraan of koelmiddelbron niet waar de
waterdruk niet geregulariseerd is.
• Stel Nooit antivriesmiddel of organische
oplosmiddelen in deel van het instrument voor.
Organische oplosmiddelen zullen onherstelbare
schade aan de eenheid veroorzaken!
• Bedien niet met buertemperaturen boven het
maximum speciceerde technische specicaties.
Oververhittend zal onherstelbare schade aan de
eenheid veroorzaken!
Tärkeää Tietoa – Finnish
• Jos tätä varusteita käytetään tavassa ei määritetty
Hoefer, Inc. suojelu ehkäisty varusteille saattaa olla
avuton.
• Tämä väline suunnitellaan sisälaboratoriokäytölle
vain.
• Vain lisävarusteet ja osat hyväksyivät tai toimitti
Hoefer, Inc. oheen ää voi käyttää käyttämiselle,
valvoalle, ja servicing tämä tuote.
• Vain käyttää käyttöjännitettä joka on CE merkitsi
tai turvallisuus joka on todistanut aidoksi ohi
joka on kansallisesti tunnustettnut testaaminen
laboratoriota.
• Turvallisuuskansi täytyy olla paikallaan
ennen yhdistäminen käyttöjännitelyijyjä
käyttöjännitteeseen.
• Kiertää kaikki käyttöjännitevalvonnat ja irrottaa
valtalyijyt ennen poistaminen turvallisuuskantta.
• Kiertää vain vesi tai 50/50 vesi/ethyleneä glycol
siinä tapauksessa varustetun lämmönvaihtimen
läpi. Älä yhdistä lämmönvaihdinta
vesinapautukseen eikä jäähdytysnestelähteeseen,
missä vesipaine on unregulated.
• Pakkasneste eikä orgaaninen liuotin välineen
osassa ei esitele Koskaan. Orgaaniset liuottimet
aiheuttavat korvaamattoman vahingon yksikköön!
• Ei käytä puskuria yllä olevia lämpötiloja
enintään määritetyillä teknisillä täsmennyksillä.
Ylikuumeneminen aiheuttaa korvaamattoman
vahingon yksikköön!
Information Importante – French
• Si cet équipement est utilisé dans une manière pas
spécié par Hoefer, Inc. la protection fourni par
l’équipement pourrait être diminuée.
• Cet instrument est conçu pour l’usage de
laboratoire intérieur seulement.
• Seulement les accessoires et les parties ont
approuvé ou ont fourni par Hoefer, Inc. pourrait
être utilisé pour fonctionner, maintenir, et
entretenir ce produit.
• utilise Seulement une alimentation qui est
CET a marqué ou la sécurité certié par un
nationalement reconnu essayant le laboratoire.
• Le couvercle de sécurité doit être à sa place avant
connecter l’alimentation mene à une alimentation.
• Tourner tous contrôles d’alimentation de et
débrancher les avances de pouvoir avant enlever le
couvercle de sécurité.
• Circuler seulement de l’eau ou 50/50 glycol d’eau/
éthylène par l’exchanger de chaleur si si équipé. Ne
pas connecter l’exchanger de chaleur à un robinet
d’eau ou à la source d’agent de refroidissement où
la pression d’eau est non régulée.
• Ne Jamais introduire d’antigel ou du dissolvant
organique dans n’importe quelle partie de
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l’instrument. Les dissolvants organiques causeront
des dommages irréparables à l’unité!
• Ne pas fonctionner avec les températures de
tampon au-dessus du maximum a spécié des
spécications techniques. La surchaue causera
des dommages irréparables à l’unité !
Wichtige Informationen – German
• Wenn diese Ausrüstung gewissermaßen nicht
angegeben durch Hoefer, Inc. verwendet wird,
kann der durch die Ausrüstung zur Verfügung
gestellte Schutz verschlechtert werden.
• Dieses Instrument wird für den
Innenlaborgebrauch nur dafür entworfen.
• Nur Zusätze und Teile genehmigten oder lieferten
durch Hoefer, Inc. kann für das Funktionieren,
das Aufrechterhalten, und die Wartung dieses
Produktes verwendet werden.
• Verwenden Sie nur eine Energieversorgung,
die CE gekennzeichnet oder durch ein national
anerkanntes Probelaboratorium bescheinigte
Sicherheit ist.
• Der Sicherheitsdeckel muss im Platz vor dem
Anschließen der Energieversorgung sein führt zu
einer Energieversorgung.
• Alle Energieversorgungssteuerungen abdrehen
und die Macht trennen führt vor dem Entfernen
des Sicherheitsdeckels.
• Nur Wasser oder 50/50 Glykol des Wassers/
Äthylens durch den Wärmeaustauscher, wenn so
ausgestattet, in Umlauf setzen. Verbinden Sie den
Wärmeaustauscher mit einem Wasserklaps oder
jeder Kühlmittel-Quelle nicht, wo der Wasserdruck
ungeregelt wird.
• Führen Sie nie Frostschutzmittel oder jedes
organische Lösungsmittel in jeden Teil des
Instrumentes ein. Organische Lösungsmittel
werden nicht wiedergutzumachenden Schaden
der Einheit verursachen!
• Mit Puertemperaturen über angegebenen
technischen Spezizierungen des Maximums
nicht funktionieren. Die Überhitzung wird nicht
wiedergutzumachenden Schaden der Einheit
verursachen!
Informazioni Importanti – Italian
• Se quest’apparecchiatura è usata in un modo
specicato da Hoefer, Inc. la protezione fornito
dall’apparecchiatura potrebbe essere indebolita.
• Questo strumento è disegnato per l’uso di
laboratorio interno solo.
• Solo gli accessori e le parti hanno approvato o
hanno fornito da Hoefer, Inc. potrebbe essere
usato per operare, per mantenere, e per revisionare
questo prodotto.
• usa Solo un alimentatore che è CE ha marcato
o la sicurezza certicato da un nazionalmente
riconosciuto testando il laboratorio.
• Il coperchio di sicurezza deve essere nel luogo
prima di collegare i piombi di alimentatore a un
alimentatore.
• Spegne tutto i controlli di alimentatore e
disinserisce i piombi di potere prima di togliere il
coperchio di sicurezza.
• Circola solo l’acqua o 50/50 glicole di acqua/
etilene attraverso lo scambiatore di calore se
così equipaggiato. Non collegare lo scambiatore
di calore a un rubinetto di acqua o qualunque
fonte di refrigerante dove la pressione di acqua è
sregolata.
• Non introduce mai l’antigelo o qualunque solvente
organico in qualunque parte dello strumento. I
solventi organici causeranno il danno irreparabile
all’unità!
• Non opera con le temperature di tampone al di
sopra del massimo ha specicato le descrizioni
tecniche. Il surriscaldamento causerà il danno
irreparabile all’unità!
Viktig Informasjon – Norwegian
• Hvis dette utstyret blir brukt i en måte ikke
spesisert ved Hoefer, Inc. beskyttelsen som ha
blitt git av utstyret kan bli svekket.
• Dette instrumentet er utformet for innendørs
laboratoriumbruk bare.
• Bare tilbehør og deler godkjente eller forsynte ved
Hoefer, Inc. kan bli brukt for drive, vedlikeholde, og
betjene dette produktet.
• bruker Bare en kraftforsyning som er CE merket
eller sikkerhet som ha blitt sertisert av et som
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nasjonalt ha blitt anerkjent prøver laboratorium.
• Sikkerheten lokket må være på plass før forbinding
kraftforsyningene blyene til en kraftforsyning.
• Vender all kraftforsyningsstyring av og frakopler
kreftene blyene før erning sikkerheten lokket.
• Sirkulerer bare vann eller 50/50 vann/ethylene
glykol gjennom oppvarmingen veksleren i så fall
utstyrer. Ikke forbind oppvarmingen veksleren
til en vanntapp eller noe kjølemiddelkilde hvor
vannet trykket er unregulated.
• Introduserer Aldri antifreeze eller noe organisk
løsemiddel inn i noe del av instrumentet.
Organiske løsemiddler vil forårsake irreparabel
skade på enheten !
• Driver med buertemperaturer over maksimum
ikke spesiserte teknisk spesikasjoner. Å
overoppheting vil forårsake irreparabel skade på
enheten !
Wazne Informacje – Polish
• Jeżeli ten sprzęt jest wykorz ystywany w sposób
nie określone przez Hoefer, Inc. do ochrony
przewidzianej przez urządzenie może zostać
obniżony.
• Instrument ten jest przeznaczony do użytku w
laboratoriach kryty tylko.
• Tylko akcesoriów i części zatwierdzone lub
dostarczone przez Hoefer, Inc. mogą być
wykorzystane do eksploatacji, utrzymania i obsługi
tego produktu.
• korzystać jedynie zasilacza że jest noszące
oznakowanie CE lub bezpieczeństwa
uwierzytelnione przez uznane na poziomie
krajowym laboratorium badawcze.
• Bezpieczeństwo lid musi być w miejsce przed
podłączeniem zasilania prowadzi do zasilania.
• Zaś wszystkie źródła zasilania urządzenia sterujące
o i odłączyć moc prowadzi przed odbiorem
bezpieczeństwa lid.
• Krążą tylko wody lub wody 50/50/ethylene glycol
wymiennik ciepła poprzez jeśli tak wyposażone.
Nie należy połączyć wymiennik ciepła woda z
kranu lub jakimkolwiek chłodziwo źródła, jeżeli
ciśnienie wody jest nieuregulowanych.
• Nigdy nie wprowadzać rozpuszczalnika
organicznego przeciw zamarzaniu lub
jakichkolwiek na dowolną część dokumentu.
Rozpuszczalniki organiczne spowoduje
nieodwracalne szkody dla jednostki!
• Nie działają w buforze temperatury powyżej
maksymalnego określone specykacje techniczne.
Przegrzania spowoduje nieodwracalne szkody dla
jednostki!
Informações Importantes –
Portuguese
• Se este equipamento é usado numa maneira
não especicada por Hoefer, Inc. que a
protecção fornecida pelo equipamento pode ser
comprometida.
• Este instrumento é projectado para uso de interior
de laboratório só.
• Só acessórios e partes aprovaram ou forneceu por
Hoefer, Inc. pode ser usada para operar, manter, e
servicing este produto.
• Só usa um estoque de poder que é CE marcou
ou segurança registrada por um nacionalmente
reconhecido testando laboratório.
• A tampa de segurança deve estar em lugar antes
de ligar o estoque de poder leva a um estoque de
poder.
• Desliga todos controlos de estoque de poder e
desconecta os chumbos de poder antes de retirar a
tampa de segurança.
• Circulam só água ou 50/50 glicol de água/ethylene
pelo exchanger de calor se for assim equiparam.
Não ligue o exchanger de calor a uma torneira de
água nem qualquer fonte de refrigerante onde a
pressão de água é não regulado.
• Nunca introduz anticongelante nem qualquer
orgânico solvente em qualquer parte do
instrumento. Orgânico solvente causará agressão
irreparável à unidade!
• Não opera com temperaturas de buer acima
do máximo especicou especicações técnicas.
Superaquecer causará agressão irreparável à
unidade!
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•
Información Importante – Spanish
• Si este equipo es utilizado en una manera no
especicado por Hoefer, Inc. la protección
proporcionado por el equipo puede ser dañada.
• Este instrumento es diseñado para el uso interior
del laboratorio sólo.
• Sólo accesorios y partes aprobaron o suministraron
por Hoefer, Inc. puede ser utilizado para operar,
para mantener, y para atender a este producto.
• Sólo utiliza una alimentación que es CE marcó o
la seguridad certicada por un nacionalmente
reconocido probando el laboratorio.
• La tapa de la seguridad debe estar en el lugar
antes de conectar la alimentación lleva a una
alimentación.
• Apaga todos controles de alimentación y
desconecta los plomos del poder antes de quitar la
tapa de la seguridad.
• Circula sólo agua o 50/50 glicol de agua/etileno
por el intercambiador de calor si ése es el caso
equiparon. No conecte el intercambiador de calor a
un toque de la agua ni cualquier fuente del líquido
refrigerante donde la presión del agua está libre.
• Nunca introduce anticongelante ni algún solvente
orgánico en cualquier parte del instrumento. Los
solventes orgánicos causarán daño irreparable a
la unidad!
• No opera con temperaturas de búfer encima del
máximo especicó especicaciones técnicas.
Recalentar causará daño irreparable a la unidad!
• Säkerheten locket måste vara på platsen före
koppla kraften tillgången blyen till en kraft tillgång.
• Vänder sig alla kraft tillgång kontroller av och
kopplar bort kraften blyen före ytta säkerheten
locket.
• Cirkulerar bara vatten eller 50/50 vatten/ethylene
glycol genom värmen exchanger i så utrustad fall.
Inte kopplar värmen exchanger till en vatten kran
eller något kylmedel källa där vattnet trycket är
unregulated.
• Inför aldrig kylvätska eller något organiska
lösningsmedel in i någon del av instrumentet.
Organiskt lösningsmedel ska orsaka irreparable
skada till enheten!
• Använd inte med buert temperaturer över
det högsta angivna tekniska specikationerna.
Överhettning skulle orsaka irreparabla skador på
enheten!
Viktig Information – Swedish
• om denna utrustning används i ett sätt som
inte har speciceras av Hoefer, Inc. skyddet
tillhandahöll vid utrustningen kan skadas.
• Detta instrument formges för
inomhuslaboratorium användning bara.
• Bara medhjälpare och delar godkände eller
levererade vid Hoefer, Inc. kan användas för
fungera, underhålla, och servicing denna produkt.
• använder bara en kraft tillgång som är CE
markerade eller säkerhet intygade vid en nationellt
erkänd testande laboratorium.
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Waste Electrical and Electronic Equipment (WEEE)
English
French
German
Italian
Spanish
This symbol indicates that the waste of electrical and
electronic equipment must not be disposed as unsorted
municipal waste and must be collected separately. Please
contact an authorized representative of the manufacturer
for information concerning the decommissioning of your
equipment.
Ce symbole indique que les déchets relatifs à l’équipement
électrique et électronique ne doivent pas être jetés comme
les ordures ménagères non-triées et doivent être collectés
séparément. Contactez un représentant agréé du fabricant
pour obtenir des informations sur la mise au rebut de votre
équipement.
Dieses Symbol kennzeichnet elektrische und elektronische
Geräte, die nicht mit dem gewöhnlichen, unsortierten
Hausmüll entsorgt werden dürfen, sondern separat
behandelt werden müssen. Bitte nehmen Sie Kontakt mit
einem autorisierten Beauftragten des Herstellers auf, um
Informationen hinsichtlich der Entsorgung Ihres Gerätes zu
erhalten.
Questo simbolo indica che i rifiuti derivanti da
apparecchiature elettriche ed elettroniche non devono essere
smaltiti come rifiuti municipali indifferenziati e devono invece
essere raccolti separatamente. Per informazioni relative alle
modalità di smantellamento delle apparecchiature fuori uso,
contattare un rappresentante autorizzato del fabbricante.
Este símbolo indica que el equipo eléctrico y electrónico no
debe tirarse con los desechos domésticos y debe tratarse por
separado. Contacte con el representante local del fabricante
para obtener más información sobre la forma de desechar el
equipo.
Swedish
Denna symbol anger att elektriska och elektroniska
utrustningar inte får avyttras som osorterat hushållsavfall och
måste samlas in separat. Var god kontakta en auktoriserad
tillverkarrepresentant för information angående avyttring av
utrustningen.
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1. Gel Electrophoresis Unit Function and Description
The Hoefer™ SE250 small format vertical slab
gel unit is intended for rapid electrophoresis of
protein or nucleic acid samples. Most samples
can be run in as little as 45 minutes, and only a
minimal amount of sample is required.
The SE250 accommodates one or two 10 × 8 cm
gel sandwiches. The upper buffer chamber is
formed when the notched side of a gel sandwich
is sealed against the silicone rubber gasket.
The upper buffer chamber core serves as a heat
exchanger if cooling is required. The core is
hollow and equipped with ports on either side
for coolant circulation.
Unpacking
Unwrap all packages carefully and compare
contents with the packing list, making sure all
items arrived. If any part is missing, contact
your local sales office. Inspect all components
for damage that may have occurred while the
unit was in transit. If any part appears damaged,
contact the carrier immediately. Be sure to keep
all packing material for damage claims or to use
should it become necessary to return the unit.
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2. Specifications
Gel plate size 10 × 8 cm
Approximate gel size 8 × 7 cm
Max. wattage 12 W
Max. voltage 500 V
Max. amperage 500 mA
Max. temperature 45 °C
Environmental Indoor use: 4– 40 °C
operating conditions Humidity up to 80%
Altitude up to 2000 m
Installation category II
Pollution degree II
Dimensions (w × h × d) 16.5 × 16 × 16 cm
(6.5 × 6.3 × 6.3 in.)
Product certifications EN 61010-1, UL 61010A-1,
CSA C22.2 1010.1,
CE Certified
This declaration of conformity is only valid for the
instrument when it is:
• used in laboratory locations,
• used as delivered from Hoefer, Inc. except for
alterations described in the user manual, and
• connected to other CE-labeled instruments or
products recommended or approved by Hoefer, Inc.
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Fig 1. Main components
SE250 electrophoresis unit.
Included but not shown:
Glass plates
Notched alumina plates
Gel seal, 1/4 oz.
Well-locating decal
Color-coded
leads (2)
SE6056-HV
Safety lid
SE256
Required but not included:
Power supply with a
minimum rating: 250 V,
50 mA, constant current
or constant voltage.
Upper buffer
chamber core
SE254B
Foam gasket
SE208
Positioning
tabs
Spring clamps (4)
SE252
Coolant port (2)
Lower buffer
chamber
SE255
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Note: All electrophoresis
accessories and kits are listed in
the the ordering section.
Note: Inspect glass plates
for chipped edges. Use only
unchipped plates to prevent
leaking.
Notched plate
3. Operating Instructions
3.1 Prepare the gel sandwich
Both precast and self-cast gels can be run in the
SE250 units. This unit accepts gels in 10 × 8 cm
plates, which can be cast in a Hoefer SE215,
SE245, or SE275 gel caster.
Each unit includes notched alumina plates and
rectangular glass plates. If casting your own
polyacrylamide gels, we recommend using a
notched alumina ceramic back plate because it
transfers heat 40 times more rapidly than glass.
For applications that are not heat sensitive, a
notched glass plate is available.
Before loading gels into the electrophoresis unit,
the separating gel should already be completely
polymerized. Clean away any gel adhering to
the notched alumina back plate. The stacking
gel (if applicable) can be cast in place on the
electrophoresis unit. Load liquid samples after
the gel sandwich is installed.
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Important! Use only water or
water and ≤50% ethylene
glycol as a coolant. Do not use
a commercial antifreeze or any
alcohol-based mixture.
Note: If the cooling option is
used frequently, it is convenient
to attach QuickFit connectors to
the tubing.
The valves in these fittings
prevent coolant spillage.
3.2 Prepare the unit
1
To disassemble a fully assembled unit: Remove the
safety lid by pressing on the handle at the top of the
upper buffer chamber core while lifting the lid by the
bottom edges. Empty all buffer chambers and remove
any gel sandwiches. Then depress both release tabs
and lift out the upper buffer chamber core.
2
Rinse the instrument before each use. Before using the
first time, disassemble the unit completely and wash
with a dilute solution of a laboratory detergent and
thoroughly rinse with water and distilled water.
3
Check the gasket. Periodically remove the gray silicone
rubber gasket from the core. Inspect for nicks and
wear. If the gasket appears to be intact, apply a light
film of Gel seal, and replace it in the groove. Avoid
stretching the gasket by laying it onto the groove and
pressing it into place.
4
Optional cooling
Circulating pressure must not exceed 0.8 bar (12 psi)
above ambient pressure. Do not connect the cooling core
to an unregulated coolant source such as a water tap.
Connect the cooling core to a circulator bath such
as the RCB20-PLUS. Slide hose clamps (4 total)
onto each end of two lengths of 8 mm (5/16") vinyl
or silicone tubing. Attach one end of each length of
tubing to a cooling core port. Attach the free ends of
each length of tubing to the circulator bath ports; one
to the inlet and the other to the outlet. Secure the
connections with the hose clamps.
p5
•
Fig 2. Core installation
and removal.
5
Install the upper buffer chamber core. First steady the
lower chamber with one hand and then hold the core
with the other hand, position it on the positioning
tabs, and press down, listening for the core to snap
into place. (Alternatively, depress both release tabs at
either side, position the core on the positioning tabs,
press into place, and release the tabs. Check that the
core is secure.)
Handle
To remove the core,
depress both release
tabs and lift.
To install the core, position it over
the positioning tabs. Then either
press down, listening for the core
to snap into place
—OR—
depress both release tabs, set the
core in place, and release.
p6
•
Release tabs (2)
Coolant port (2)
Fig 3. Gel sandwich installation.
A 10 × 8 cm gel sandwich fits
flush with the bottom of the upper
buffer chamber core.
3.3 Place the gel sandwich
1
Rinse away the overlay with distilled water and drain
any excess water.
2
If installing a self-cast or precast 10 × 8 cm gel
sandwich, orient the sandwich so that the notched
plate faces the gasket, notches at the top. Set the
bottom of the sandwich on the supporting ledges in
the bottom of the lower chamber and center the plate
so that the gasket seals both sides (Fig 3).
p7
•
Clamp the sandwich in place
1
Lightly press the sandwich against the gasket and
secure it to the core with one spring clamp on each
side. Position the jaw so that the shorter rounded jaw
edge fits into the core groove and the longer edge sits
on the glass plate. (Proper positioning is important to
achieve a seal and to minimize glass breakage.) Slide
the clamps down to the stop.
Fig 4. Securing the gel
sandwich onto the upper buffer
chamber core.
Each sandwich requires two
clamps. The rounded edge of the
short jaw on the clamp fits into
the groove behind the gasket, and
the long jaw presses on the glass
plate over the spacer.
2
Repeat step 1 for the second sandwich, or, if running
only one gel, clamp a plain glass plate on the unused
side of the core to prevent a possible short circuit
with the unused electrode. (Do not fill this chamber
with buffer if no gel sandwich is in place.)
Fit short jaw of clamp
into the groove
Cooling is optional:
Attach tubing to ports on
both sides of the core before
attaching gel sandwiches.
Circulate coolant.
p8
•
Note: Stacking gel resolution is
optimal when poured just before
electrophoresis.
3.4 Sample preparation and loading
1
If wells are already in place, skip to step 2.
If applicable, cast the stacking gel in the unit.
Calculate the stacking gel monomer solution volume:
measure the distance, in cm, from the top of the
resolving gel to the notch in the alumina plate. (This
should be at least 2 cm — more if the sample depth
in the well is unusually high.) Multiply this distance
by the gel width (8.3 cm) and the gel thickness (cm).
This product is the required volume in ml.
Deaerate the stacking gel monomer solution, add
catalyst and initiator and then pour. Use a pipette to
deliver the solution into one corner of the plate, taking
care not to trap any bubbles. Insert a comb (at a
slight angle to prevent trapping air) into the sandwich,
allowing the comb sides to rest on the spacers.
Overlay each gel with a thin layer of water-saturated
n-butanol, water, or diluted gel buffer to prevent gel
exposure to oxygen. Slowly deliver the overlay solution
from a glass syringe fitted with a 22-gauge needle.
Apply the solution near the spacer at the side of
the sandwich and allow it to flow across the surface
unaided. Allow a minimum of one hour for the gel
to polymerize.
2
Prepare the sample. Increase liquid sample density
with 10% glycerol or sucrose. Add a tracking dye such
as phenol red or bromophenol blue.
For SDS protein gels, use 2X treatment buffer to
denature both liquid and dry samples in a test tube.
To liquid protein solutions, add an equal volume
of 2X buffer. To dry protein samples, add equal
volumes of buffer and ddH2O to achieve the desired
concentration. Heat the tube in boiling water for 90
seconds, then chill it in ice until ready to use. Treated
samples can be stored frozen for future runs. (Store at
-40 °C to -80 °C.)
p9
•
Note: The amount of protein
sample added to each well
depends on both the sensitivity
of the staining method and
the distribution of protein
among separate bands. With
Coomassie™ Blue, it is possible
to detect 1 µg in a single band;
with the more sensitive silver
stains, it is possible to detect as
little as 10 ng.
3
To aid in loading samples, wet the well-locating decal
and apply it to the front of the glass plate so that the
appropriate edge outlines the sample wells.
Note: The side wells for standards of a preparative
comb correspond to the outer-most wells formed by
the 10-well comb.
4
Fill the sample wells and each upper buffer chamber
that will be used with running buffer. One upper
buffer chamber holds approximately 75 mL.
5
Underlay the sample into the wells using a fine-tipped
microsyringe. The width of the wells depends on the
number of wells per comb. If the comb has fewer
wells, they are wider, and require more volume to
raise the level 1 mm, as shown in the following table.
Volume of sample (µL) per 1 mm depth
no. of comb thickness (mm)
wells 0.75 1.0 1.5
5 9.5 12.7 19.1
9 5.8
10 3.6 4.8 7.2
15 2.2 2.9 4.4
18 2.9
•
p10
Note: If using precast gels,
check that the lower gel/buffer
contact surface is exposed.
Important! Do not use antifreeze
or any alcohol-based mixture,
as these will irreparably damage
the core.
3.5 Final assembly
1
Fill the lower buffer chamber with running buffer. The
SE250 holds about 150 mL. Check that the lower
electrode (running along the bottom of the the upper
buffer chamber core) is completely submerged.
2
Place the safety lid on the unit.
3
Plug the color-coded leads into the jacks of an
approved power supply such as the PS300B. The red
lead plugs into the red output jack, and the black
lead plugs into the black output jack.
4
Optional cooling: Begin circulating cold water or a
chilled 50/50 water/ethylene glycol solution.
•
p11
3.6 Running the gel
Gels may be run at either constant current or
constant voltage. A constant current setting is
traditionally used with a discontinuous buffer
system so that the rate of electrophoretic
migration remains unchanged throughout the
run. Under these conditions, voltage increases
as the run proceeds. A lower current setting
is recommended for higher resolution. Precast
gels are run under the same current and voltage
conditions as self-cast gels.
Important! After initial
monitoring, do not leave the
unit unattended for more than
45 minutes before checking the
progress of the the bands and
the buffer level.
It takes about one hour to run two 7 cm ×
0.75 mm Laemmli gels at 40 mA (20 mA per gel,
constant current). Check band progress after
5 minutes, and again after half an hour, keeping
an eye on the position of the tracking dye. The
run is complete when the tracking dye reaches
the bottom of the gel. Watch the buffer level
in the upper buffer chamber and, if necessary,
replenish it before it falls below the level of the
notched plate. (A small volume of buffer may
leak past a chipped plate or nicked gasket, or it
may wick out through the gel.)
•
p12
Important! Always disconnect
the high voltage leads from the
power supply before removing
the lid from the unit.
After the run
1
Once the tracking dye reaches the bottom of the gel,
turn off the power supply, disconnect the leads, and
remove the safety lid.
2
If coolant is circulating, stop the flow and disconnect
the fittings or tubing.
3
Remove the core assembly with gels attached by
squeezing the release tabs and lift out the core
assembly.
4
Pour out the buffer by inverting the core assembly,
then remove both clamps, and lift away gel
sandwich(es) from the upper buffer chamber core.
5
Gently loosen and then slide away both spacers. Slip an
extra spacer or a Hoefer Wonder Wedge into the bottom
edge (to prevent breaking the ears of the notched
plates) and separate the plates. The gel usually adheres
to the alumina plate. Carefully lift the gel from the plate
and lay it into a tray of stain or fixative.
•
p13
4. Care and Maintenance
• Do not autoclave or heat any part above 45 °C.
• Do not use organic solvents, abrasives, strong
cleaning solutions, or strong acids or bases to
clean the chambers.
• Immediately after each use, rinse the unit with
water and then rinse thoroughly with distilled
water. Handle the upper buffer chamber core
with care to prevent damage to the banana
plugs. Allow to air dry.
• Clean glass and alumina plates and spacers
with a dilute solution of a laboratory cleanser
such as RBS-35™, then rinse thoroughly with
tap and distilled water. Glass plates can also be
treated with (but not stored in) acid cleaning
solutions.
•
p14
5. Troubleshooting
problem solution
Smile effect on the buffer front To reduce the running temperature:
• Circulate coolant through the upper buffer
chamber core.
• Prechill the buffer.
• Decrease the current or voltage setting. (10 mA per
0.75 mm gel, 15 mA per 1.5 mm thick gel.)
• Run the gel in the cold room.
Protein streaks vertically • Centrifuge or filter sample before loading to remove
particulates.
• Dialyze or desalt the sample.
Unusually slow (or fast) run Adjust the solutions:
• Check recipes, gel concentrations, solutions, and
dilutions. (For instance, do not use Tris-HCl instead
of Tris.)
• If the required pH of a solution is exceeded, do not
back-titrate. Prepare fresh buffer.
• Dispose of older acrylamide solutions and use only
stock of the highest quality.
• Only use freshly deionized urea.
Adjust the voltage or current settings:
• To increase or decrease the migration rate, adjust
the voltage or current by 25–50%.
Bands are skewed or distorted Check gel preparation and polymerization:
• Degas the stacking gel solution and avoid trapping
air bubbles under the comb teeth.
• Overlay the running gel with water-saturated
n-butanol before polymerization begins to avoid
forming an uneven gel surface.
Check sample preparation:
• Dialyze or desalt the sample.
• Centrifuge or filter sample before loading to remove
particulates.
•
p15
problem solution
Stained sample collects: Near the buffer front:
• Protein is not sufficiently restricted by the resolving
gel; increase the % T.
Near the top of the gel when the buffer front has
reached the bottom:
• The gel pore size is too small. Decrease the % T of
the resolving gel.
• The protein has precipitated. Heat the sample at a
lower temperature (70 °C or less) for 1–2 minutes.
Poor band resolution • Use only the highest quality reagents.
• Conduct the separation at a lower current or
voltage setting.
• Dialyze or desalt the sample.
• Reduce the sample volume or concentration.
• Only use freshly deionized urea.
• Improve dissociation of subunits by heating sample
in SDS sample buffer 1–2 minutes at 100 °C.
• Add more mercaptoethanol or dithiothreitol; check
sample treatment.
• Only use gels that were recently prepared.
• Check pH values of the separating and stacking gel
solutions. Do not back-titrate buffers.
Sample preparation:
• Heat samples for no more than 1–2 minutes at
100 °C. Store on ice after heating.
• Store sample on ice before it is denatured.
• Add protease inhibitors if necessary to prevent
proteolytic degradation of sample.
• Store samples to be frozen in aliquots to prevent
repeated freezing and thawing. (Store at -40 °C to
-80 °C.)
Bromophenol blue doesn’t sharpen • Pour a taller stacking gel. (For best results, allow a
into a concentrated zone in the • stacking gel height of 2.5 times the height of the
stacking gel • sample in the well.)
• Dispose of outdated acrylamide solutions and use
only the highest grade of acrylamide.
• When preparing samples, avoid using solutions with
a high sodium or potassium concentration.
•
p16
SE250 results:
Lane 1: SDS-6H, high MW
standard mixture, Sigma
Lane 2: SDS-7 Dalton Mark
VII-L™, Sigma (10 µl per lane)
Gel
12% SDS PAGE
Stained with Coomassie Blue
Running conditions
20 mA, one hour
™
Appendix
The following Laemmli system is slightly modified
for use with the mini-vertical units. The Laemmli
system is the most common electrophoresis
protocol for SDS-denatured proteins. The leading
ion in this discontinuous buffer system is chloride
and the trailing ion is glycine. Accordingly, the
resolving gel and the stacking gel contain Tris-Cl
buffers (of different concentration and pH), and
the electrophoresis buffer contains Tris-glycine.
All buffers contain 0.1% SDS.
Polyacrylamide gel composition is indicated by
two different percentages:
% T = total acrylamide = g (acryl + bis) × 100
100 mL
% C = crosslinker = g (bis) × 100
g (acryl + bis)
The total percent of acrylamide (% T) in the
separating gel, which can range from 5 to
20%, determines the pore size. Commonly,
the amount of crosslinker used (% C) is 2.6%.
In the following example system, the resolving
gel composition is 10% T, 2.6% C, which
results in a medium pore size. The stacking gel
composition is 4% T, 2.6% C. The % T in the
stacking gel is lower because a larger pore size is
required.
•
p17
Final concentrations
separating gel stacking gel electrophoresis
Acrylamide concentration 10% T*, 2.6% C 4% T, 2.6% C
Tris-Cl 0.375 M 0.125 M
Tris-glycine 0.025 M Tris base
0.192 M glycine
pH 8.8 6.8 ~8.3
SDS 0.1% 0.1% 0.1%
Ammonium persulfate (APS) 0.05% w/v 0.05–0.1% w/v
TEMED† 0.05% v/v 0.05–0.1% v/v
* To achieve any other desired final concentration, adjust the acrylamide stock and water volumes.
†Tetramethylethylenediamine
buffer
•
p18
Bibliography
Adams, L.D. and Gallagher, S.R., Two-Dimensional
Gel Electrophoresis Using the O’Farrell System.
Current Protocols in Molecular Biology,
10.4.1–10.4.13 (1992).
Gallagher, S.R., and Smith, J.A., Electrophoretic
separation of proteins. Current Protocols in
Molecular Biology. (F.A. Ausubel, R. Brent, R.E.
Kingston, D.D. Moore, J.G. Seidman, J.A. Smith,
and K. Struhl, eds.) 10.2.1–10.2.21 (1991).
Laemmli, U.K., Cleavage of structural proteins during
the assembly of the head of bacteriophage T.
Nature. 227, 680–685 (1970).
Matsudaira, P.T. and Burgess, D.R., SDS microslab
linear gradient polyacrylamide gel electrophoresis.
Anal. Biochem. 87, 386–396 (1978).
Reisfeld, R.A., et al. Acidic buffer system for
resolution of cationic proteins. Nature. 195, 281
(1962).
Sasse, J., and Gallagher, S.R., Staining proteins in
gels. Current Protocols in Molecular Biology. (F.A.
Ausubel, R. Brent, R.E. Kingston, D.D. Moore,
J.G. Seidman, J.A. Smith, and K. Struhl, eds.)
10.6.1–10.6.8 (1991).
Towbin, H., et al. Electrophoretic transfer of proteins
from polyacrylamide gels to nitrocellulose:
procedure and some applications. Proc. Natl. Acad.
Sci. USA. 76, 4350–4353 (1979).
Weber, K., and Osborn, M., The reliability of
molecular weight determinators by dodecyl sulfatepolyacrylamide gel electrophoresis. J. Biol. Chem.
224, 4406–4412 (1969).
•
p19
Ordering Information
product quantity code number
Hoefer SE250 Mini-gel System 1 SE250-10A-.75
Mini-Vertical Unit for 2 slab gels, Complete
Includes: basic unit, 10 glass plates (10 × 8 cm),
2 alumina notched plates, well-locating decal,
SE245 Dual Gel Caster, 2 each 0.75-mm thick 10-well combs
and 0.75-mm thick spacer sets.
Electrophoresis Unit Replacement Parts
Foam gasket, 4.5 mm × 61 cm 1 SE208
Upper buffer chamber for SE250 1 SE254B
Lower buffer chamber for SE250 1 SE255
Lid with cables for SE250 1 SE256
Wonder Wedge plate separation tool 1 SE1514
High voltage safety lead set 1 SE6056-HV
Gel Seal (0.25 oz.) 1 SE6070
Spring clamps, for SE250 and gel casters 4 SE252
Well locating label 2 SE212
Glass and Alumina Plates
10 × 8 cm
Notched alumina plates 10 SE202N-10
Rectangular glass plates 10 SE202P-10
•
p20
Spacers
thickness (mm) length (cm) quantity code number
0.75 8 2 SE2119T-2-.75
1.00 8 2 SE2119T-2-1.0
1.50 8 2 SE2119T-2-1.5
Combs
wells thickness (mm) width (mm) quantity code number
5 0.75 13.0 1 SE211A-5-.75
5 1.00 13.0 1 SE211A-5-1.0
5 1.50 13.0 1 SE211A-5-1.5
9a 1.00 5.8 1 SE211A-9-1.0
10 0.75 4.8 1 SE211A-10-.75
10 1.00 4.8 1 SE211A-10-1.0
10 1.50 4.8 1 SE211A-10-1.5
12 1.00 4.75 1 SE211A-12-1.0
15 0.75 2.9 1 SE211A-15-.75
15 1.00 2.9 1 SE211A-15-1.0
15 1.50 2.9 1 SE211A-15-1.5
18a 1.00 2.9 1 SE211A-18-1.0
1/1b 0.75 68/5 1 SE211A-R-.75
1/1b 1.00 68/5 1 SE211A-R-1.0
1/1b 1.50 68/5 1 SE211A-R-1.5
a
Microtiter spacing
b
Preparative/reference well
•
p21
product quantity code number
Gel Casters
For 1 or 2 gels, 10 × 8, 10.5
Hoefer SE245 Dual Gel Caster 1 SE245
For 5 to 10 gels, 10 × 8 cm
Hoefer SE215 Multiple Gel Caster, Complete 1 SE215
Includes: 20 rectangular glass plates, 10 notched
alumina plates, 100 sheets of wax paper, space saver plate,
5 filler sheets, set of filler plugs and Spacer-Mate.
(Order combs and spacers separately.)
For 2 to 4 gels, 10 × 8 cm
Hoefer SE275 4-Gel Caster, Complete 1 SE275
Includes: 10 rectangular glass plates, 4 notched alumina
plates, 100 sheets of wax paper, space-saver plate,
5 filler sheets, Spacer-Mate and filler plugs.
(Order combs and spacers separately.)
Power Supplies
Hoefer PS300B-300V, 500 mA, 90 W 1 PS300B
Miscellaneous
Hoefer SE100 PlateMate washing and storage unit 1 SE100
TE22 Transphor Tank Unit 1 TE22
QuickFit connector, male, 3/8" 1 QFX3/8
•
p22
Hoefer, Inc.
84 October Hill Road
Holliston, MA 01746
Toll Free: 1-800-227-4750
Phone: 1-508-893-8999
Fax: 1-508-893-0176
E-mail: support@hoeferinc.com
Web: www.hoeferinc.com
Hoefer is a registered trademark
of Hoefer, Inc. Coomassie and
Tris are trademarks of ICI plc.
Dalton Mark VII-L and Sigma are
trademarks of Sigma Chemical Co.
RBS-35 is a trademark of Pierce
Chemical Co.
© 2012 Hoefer, Inc. — All rights
reserved.
Printed in the USA.
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