Hoefer SE 600 Chroma User Manual
user manual
SE 600 Chroma
Hoefer SE 600 Chroma
standard dual cooled gel electrophoresis unit
um SE600X-IM/Rev. A0/05-04
Page finder
Gel electrophoresis unit function and description
Specifications .................................................................
Important information ....................................................
Unpacking and inventory ....................................................
Operating instructions
Prepare the gel sandwich .................................................
Construct the gel sandwich & insert into caster ..................... 9
Acrylamide gels ......................................................................12
Gradient gels ..........................................................................14
Sample preparation and loading ...........................................16
Final assembly ................................................................18
Separating the sample ...................................................22
After electrophoresis .......................................................24
Care and maintenance .....................................................25
Troubleshooting .................................................................26
Bibliography .......................................................................31
Ordering information ........................................................34
Companion products .........................................................38
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Gel electrophoresis unit
function and description
The Hoefer™ SE 600 Chroma™ vertical slab gel
electrophoresis unit is intended for protein and
nucleic acid electrophoresis under commonly
used denaturing and non-denaturing conditions.
Up to 28 samples can be compared on a single
slab gel.
Applications include protein separations, nucleic
acid fractionation, and the second-dimension
separation of 2-D electrophoresis. First-dimen
sion separation of 2-D protein electrophoresis
should be performed on Immobilized pH Gra
dient Gels. The focused strips are easily trans
ferred to the second-dimension slab gel for size
separation.
The gel plates are 18 cm wide by 16 cm long.
Up to four gels can be run at one time if sand
wiches are paired into “club sandwiches.” The
heat exchanger allows buffer temperature con
trol in the lower chamber.
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Specifications
Gel plate size 18 × 16 cm (w × h)
Gel size 14 or 16 cm
(w × h)
Maximum watt 50 W
Maximum volt 1 000 V
Maximum ampere 500 mA
Maximum temperature 45 °C
Environmental
operating conditions Indoor use: 4–40 °C
Humidity up to 80%
Altitude up to 2 000 m
Installation category: II
Pollution degree: 2
Dimensions width
32 × 29 × 14 cm
(12.5 × 11.5 × 5.5 in)
Product certifications EN 61010-1, UL 61010A-1,
CSA C22.2 1010.1, CE Certified
This declaration of conformity is valid only when the
instrument 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 recom
mended or approved by Hoefer, Inc.
× 16 cm
× height × depth
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Important information
• The safety lid must be in place before connecting the power
leads to a power supply.
• Turn all power supply controls off and disconnect the power
leads before removing the safety lid.
• Circulate only water or 50/50 water/ethylene glycol through the
heat exchanger. Never introduce antifreeze or any organic solvent
into any part of the instrument. Organic solvents will cause
irreparable damage to the unit!
• Do not connect the heat exchanger to a water tap or any coolant
source where the water pressure is unregulated.
• Do not operate with buffer temperature above 45 °C. All plastic
parts are rated for 45 °C continuous duty. Circulate coolant
through the heat exchanger during electrophoresis to minimize
heating. Overheating will cause irreparable damage to the unit!
• Only accessories and parts approved or supplied by Hoefer, Inc.
may be used for operating, maintaining, and servicing this
product.
Informations importantes
• Le couvercle de sécurité doit être en place avant de brancher les
prises au générateur.
• Eteindre le générateur et débrancher les prises avant d’enlever le
couvercle de sécurité.
• Faire circuler seulement de l’eau ou 50/50 d’eau et d’éthylène
glycol dans l’échangeur vertical à cirulation d’eau. Ne jamais
utiliser d’anti-gel ou tout autre solvant organique avec cet
instrument. Les solvants organiques causeraient des dommages
irréparables à l’appareil.
• Ne pas connecter l’échangeur vertical à circulation d’eau à un
robinet ou quelque source de refroidissement dont la pression
n’est pas régulière.
• Ne pas utiliser avec un tampon à une température au dessus de
45 °C. Toutes les piéces en plastique sont prévues pour résister
à une température constante de 45 °C. Faire circuler l’eau dans
l’échangeur vertical durant l’électrophorèse pour minimiser
l’échauffement afin d’éviter des dommages irréparables à
l’instrument.
• Seulement les accessoires et piéces detachées approuvés ou
fournis par Hoefer, Inc. sont recommandés pour l’utilisation,
l’entretien et réparation de cet appareil.
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Fig 1. Main components of the
Hoefer SE 600 Chroma
(see Fig 4 for caster compo
nents).
Included but not shown:
• Gel Seal compound, 1/4 oz.
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color-coded
leads (2)
safety lid
• Spacer-Mate alignment
template
• Well-locating decal
• Glass plates (6)
• Wonder Wedge plate separa
tion tool
• Buffer dam
Complete unit also includes
spacers (4) and combs (2).
Required but not included:
• Magnetic stirrer
• Power supply with a
minimum rating of 500 V,
100 mA (constant A or V)
Optional: Circulator bath
Note: The ordering section
lists all accessories and
replacement parts.
interlock pins
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upper
buffer
chamber
with upper
electrode
heat
exchanger
with lower
electrode
lower buffer
chamber
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Unpacking and inventory
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 dam
aged, 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.
Lower buffer chamber
The lower buffer chamber is transparent,
which allows visual tracking of electrophoresis
process. The chamber is chemically resistant
to common electrophoretic buffers but not to
organic solvents or strong acids or alkali. Tem
peratures above 45 °C may cause the chamber
to warp.
Upper buffer chamber
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The upper buffer chamber is chemically resistant
to common electrophoresis buffers, but not to
organic solvents or strong acids or alkali. The
upper electrode (cathode) runs along the center
ridge and terminates at the banana plug. The
upper chamber requires 0.5–0.8 l of buffer (fill
no higher than the top of the plastic ribs).
Heat exchanger
The heat exchanger must be installed for every
use because it houses the bottom electrode
(anode), which runs along the bottom of the
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frame. When connected to a circulator bath, the
heat exchanger regulates the buffer temperature
in the lower chamber. Coolant passes through
the glass tubes, which are secured with silicone
rubber grommets. The heat exchanger connec
tor ports are 13 mm o.d. The heat exchanger is
rated to a maximum of 0.8 atmospheres above
ambient (12 psig). Connect only to coolant
sources with regulated pressure. (Do not con
nect to the water tap.)
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Safety lid
The banana plug on the heat exchanger connects
to the red lead, and the plug on the upper buffer
chamber connects into the black lead. The 4 mm
shrouded color-coded leads plug into color-coded
jacks in the power supply. Engage interlock pins
before lowering electrode connections on to
banana plugs. Always install the safety lid before
use!
Glass plates
The SE 600 Chroma accommodates 18-cm-wide
plates 16 or 8 cm long. Notched divider plates,
ordered separately, divide gel sandwiches to
form “club sandwiches” of two gels each, so up
to four gels can be run at one time.
Clamps
Two 16 cm clamps are used to secure the gel
sandwich. The clamp pressure bar, adjusted with
screws, distributes pressure evenly.
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Casting stand
The casting stand holds assembled gel sandwiches upright for casting gels. Adjustable
feet level the caster. A laminated gasket in the
bottom of each casting cradle seals the bottom
of the sandwich when it is clamped into the
stand.
Cams
Cams are used twice: first to secure the assembled sandwich in the casting stand and, second,
to attach the sandwich to the upper buffer
chamber.
Rubber gaskets
There are two sets of two gaskets: The solid
laminated gaskets fit into the bottom of the
casting stand and form the seal for casting the
gel. The slotted gaskets fit under the upper
buffer chamber and form the seal between the
upper and lower chambers. The ridges on the
upper gasket align the gasket slot to maintain
an open channel between the top of the gel and
the buffer in the upper chamber.
Spacers
Spacers determine the thickness of the gel and
are available in three thicknesses (0.75, 1, and
1.5 mm) and two widths (1 and 2 cm).
(May be ordered separately.)
Spacer-Mate alignment template
This template aligns spacers during sandwich
assembly.
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Combs
Teflon combs are available in sizes that form
10, 15, 20, or 28 wells. Most combs are avail
able in all three thicknesses: 0.75, 1, and 1.5
mm. Blank combs form a single large well, and
preparative combs include one or two reference
wells in addition to the preparative well.
(May be ordered separately.)
All blanks, preparative combs, and 10-, 15-, and
20-well combs form wells that are 25 mm deep.
The 28-well comb forms wells that are only 15
mm deep so that wells do not collapse when
the comb is removed. The sample volume held
by each well depends on the gel thickness, well
depth, and the number of wells per comb. Table
1 lists sample volumes of wells for all combs
(see page 10).
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Wonder Wedge Gel Plate Separation tool
This tool is used to disassemble gel sandwiches
and to check spacer and comb thicknesses.
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Operating instructions
Gel casting and electrophoresis procedures
follow. Included are instructions for polyacryl
amide gels (used with continuous or discon
tinuous buffer systems) and gradient gels. See
page 31
for bibliography.
Prepare the gel sandwich
Glass plates, spacers, and clamp sets are sized
so that the assembled sandwich can be easily
aligned to create the seal required first to cast the
gel and then to run it. For best results take extra
care to align all components when assembling
sandwiches. One to four gels (18
be assembled and run in the SE 600 Chroma.
Both precast gels and self-cast gels can be used.
To self-cast multiple gels, kits can be ordered
separately: The SE 615 Multiple Gel Caster Kit
holds up to 10 single gel sandwiches, and the SE
675 Gel Caster Kit holds up to four sandwiches.
(See the accompanying gel caster User Manual
for complete instructions.) To run four gels con
currently, two accessory notched divider plates
and two additional pairs of spacers are required.
× 16 cm) can
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Construct the gel sandwich and insert into
caster
Prepare the caster and clamps
Place the spirit level into the caster center and adjust the leveling
feet. Loosen all clamp screws and make space for the sandwich by
sliding the pressure plates toward the screws.
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Fig 2. Sandwich assembly.
glass plates
spacer
clamp ridges
pressure
plate
Inspect glass plates for nicks. Use
only unchipped plates to prevent
leaking.
The glass plates and
spacers must be flush
with the clamp ridges
at both top and bottom
for a good seal.
Fig 3. Club sandwich assembly.
Side clamps will accommodate two
spacers up to 1.5 mm thick.
glass plates
(at the outer sides
of the sandwich)
notched
center plate
spacers
Construct gel sandwiches
For each sandwich choose two perfectly clean, unchipped glass
plates and two spacers. Lay one plate on a flat surface, lay the
Spacer-Mate alignment template onto the plate (wide side at the
top of the plate), place a spacer along each edge, and lay the sec
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ond glass plate on top.
Secure the sandwich with clamps
Slide one clamp at a time along the sandwich sides. Finger-tighten
one screw on each clamp, set the sandwich upright on a flat
surface, and loosen the screw to align the stack. Taking great care
in alignment will ensure a good seal. Finger-tighten all screws.
Remove the Spacer-Mate.
Tip: Use the casting cradle to hold the sandwich during align
ment. Remove the laminated gasket from the cradle and, instead
of setting the sandwich upright on a flat surface, set it into the
casting cradle.
Club sandwich
A 16-cm-long, notched center-divider plate (ordered separately)
pairs two sandwiches to double the number of gels that can be
cast and run.
Assemble a club sandwich in the same manner as a regular sand
wich, except before placing the top glass plate, lay the divider plate
and a second set of spacers on the stack. Place the notch so that
it will be at the top of the gels. It is essential that the spacers and
plates align perfectly in order to seal.
Remove the sandwich and inspect the bottom to make sure that
edges are aligned flush to ensure a complete seal. Adjust if
necessary.
Optional: Apply a light film of Gel Seal compound only on the
bottom corner surfaces created by the spacers and plates if the
sandwiches tend to leak.
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