Instruction Manual
Precision Plus Protein
™
Standard Plugs, Unstained
Catalog #161-0378
Product shipped at room temperature.
Store at 4ºC upon arrival.
Products for 2-D Electrophoresis, Proteomics
Catalog # Description
161-1511 PROTEAN Plus Tris-HCl Gel, 12%, 24 cm IPG well
161-0787 Bio-Safe Coomassie Stain, 5 L
161-0449 Silver Stain Plus Kit
161-3138 SYPRO Ruby Protein Gel Stain, 5 L
161-0737 Laemmli Sample Buffer, 30 ml
161-0772 10 x Tris/Glycine/SDS, 5 L cube
163-2092 PROTEAN Plus Overlay Agarose
170-3990 Trans-Blot Plus Cell With Plate Electrodes and
162-0251 Nitrocellulose Membrane, 0.45 μm, 26.5 x 28 cm, 10 sheets
162-0252 Nitrocellulose Membrane, 0.2 μm, 26.5 x 28 cm, 10 sheets
162-0253 Supported Nitrocellulose Membrane, 0.45 μm,
162-0254 Supported Nitrocellulose Membrane, 0.2 μm,
162-0255 Immun-Blot PVDF Membrane, 25 x 28 cm, 10 sheets
162-0256 Sequi-Blot PVDF Membrane, 25 x 28 cm, 10 sheets
Super Cooling Coil
26.5 x 28 cm, 10 sheets
26.5 x 28 cm, 10 sheets
Products for Blot Detection
161-0380 StrepTactin-HRP Conjugate, 0.3 ml, 150 applications
161-0382 StrepTactin-AP Conjugate, 0.3 ml, 150 applications
170-5040 Immun-Star HRP Chemiluminescent Substrate, 500 ml
170-5012 Immun-Star AP Substrate Pack
This product is sold under license from Life Technologies Corporation,
Carlsbad, CA, for use only by the buyer of the product. The buyer is not
authorized to sell or resell this product or its components.
Bio-Rad Laboratories, Inc.
2000 Alfred Nobel Dr., Hercules,CA 94547
(510) 741-1000
4117684 Rev C
Representative
lot of Precision
Plus Protein
standard plug
on a 4–20% Tris-HCl
SDS-PAGEgel
kD
250
150
100
75
50
37
25
20
15
10
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Precision Plus Protein Unstained
Standard Plugs allow you to easily, quickly,
and cleanly load molecular weight standards
on any gel. The newest addition to the family
of Precision Plus Protein standards, these
ready-to-use standard plugs provide a convenient and reliable solution to molecular weight
determination on 2-D electrophoresis gels.
Specifically designed for use with PROTEAN
Plus precast gels and other gels without a
separate standards well, Precision Plus Protein
unstained standards are cast in 1 mm thick
agarose plugs for easy storage, handling, and
loading. Load concentrations have been
optimized for SYPRO, Ruby, Silver Stain Plus,
and Bio-Safe Coomassie staining. Precision
Plus Protein standard plugs come in an easy to
use, snap-off plastic plug casing, in packs of 2 x
12 (24 applications), with one application per
plug.
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Product Description
Precision Plus Protein standard plugs contain ten
recombinant protein bands of 10 kD, 15 kD, 20 kD,
25 kD, 37 kD, 50 kD, 75 kD, 100 kD, 150 kD, and
250 kD.The 25 kD, 50 kD, and 75 kD bands are
three times the intensity of the other bands and
serve as reference markers.
The molecular weights of Precision Plus Protein
standard plugs, are confirmed by mass spectrometry and migration in a Laemmli SDS-PAGE system.
Precision Plus Protein standard plugs contain an
integral Strep-tag sequence, enabling parallel
detection on western blots using StrepTactinHRP (catalog #161-0380) or StrepTactin-AP (catalog #161-0382).
3
Specifications
Catalog # 161-0378
Number of applications 24 plugs
Protein quantity 250 kD band, 540 ng; 150 kD band,
Plug buffer 1% agarose, 128 mM glycine,
Shelf life 3 months at 4ºC
315 ng; 100 kD band, 225 ng; 75 kD
band, 990 ng; 50 kD band, 1,100 ng;
37 kD band, 330 ng; 25 kD band,
1,200 ng; 20 kD band, 225 ng;
15 kD band, 300 ng; 10 kD band,
210 ng each*
* These values should be used only to
determine a rough approximation of
the quantity of a protein of interest.
10% (w/v) glycerol, 0.7% SDS,
62.5 mM Tris, pH 8.3, 50 mM DTT,
5 mM EDTA, 0.02% NaN
0.003% Bromophenol Blue
4
,
3
Instructions
Precision Plus Protein Standard Plugs are ready-touse 2-D electrophoresis standards. When ready to
use Precision Plus Protein standard plugs, break off
one of the ends of the plastic plug casing. For each
application, break off an individual plastic plug by
placing pressure on the flat side to break the
junction, then twisting the individual plug off.** Then
break off the attached spatula. Prepare SDS-PAGE
gels by blotting away any excess water remaining
inside the IPG well using blotting paper. Then use
the spatula to slide the plug outward and onto the
top plate of your gel. Use the spatula to push the
plug into the well next to the IPG strip. See Figures
1-8 for detailed illustrations of how to use the
Precision Plus Protein standard plugs.
The Precision Plus Protein standard plugs
have been optimized for 12% PROTEAN Plus
gels. If you are using a gel of a
different formulation, add 5% BME or 50 mM
DTT to your overlay agarose.
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Flat side of plastic plug casing
Plug Casing
Break Line
Spatula Break Line
Spatula
Recommended Loading Volumes for
Large Gel Systems
Load PAGE Application
Full plug Coomassie Blue G-250 or R-250 staining
1/3 plug Silver staining; SYPRO Ruby staining
** Note: To store unused plugs, it is critical to wrap the
open end of plastic plug casing tightly with Parafilm
before storage to maintain integrity of the plugs.
6
Fig. 1. Flat side of plastic casing with 12 Precision Plus Protein
standard plugs, unstained.
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Fig. 2. Break and twist off one end of the plastic plug casing.
8
Fig. 3. Break off an individual plastic plug from the casing by
placing pressure on the flat side to break the junction.
Study Fig. 3 and 4 carefully before proceeding.
9
Fig. 4. Complete the break by twisting off the individual plastic
plug casing.
10
Fig. 5. Snap off the attached spatula.
11
Fig. 6. Use spatula to push plug out of the plastic casing.
12
Fig. 7. Use the spatula to slide the plug outward and on to the
top plate of your gel.
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Fig. 8. Slide plug all the way down the bottom of the well next
to IPG strip, then add agarose overlay.
Coomassie is a trademark of Imperial Chemical Industries. Parafilm is
a trademark of American National Can Co. Strep-tag and StrepTactin
are trademarks of Institut für Bioanalytik GmbH. Strep-tag technology
for western blot detection is covered by US patent 5,506,121 and by
UK patent 2,272,698. StrepTactin is covered by German patent
application P 19641876.3. Bio-Rad Laboratories is licensed by Institut
für Bioanalytik GmbH to sell these products for research use only.
SYPRO is a trademark of Molecular Probes. Bio-Rad is licensed by
Molecular Probes to sell SYPRO products for research use only, under
US patent 5,616,502.
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