2D ELECTROPHORESIS WITH COOMASSIE BLUE
STAINING
Coomassie
blue is our most quantitative
stain as shown
in the adjacent plot of stain density > background versus total ug
rat liver cytosol loaded. for polypeptide spot 8 of MW 76,100,
pI 8.1. See
Method Validation
for detailed validation results confirming the linearity and
reproducibility of Coomassie blue staining of our large format 2D gels.
Usually 1 ug of a purified protein gives a dark spot with CB while
0.1 ug gives a faint spot. Coomassie Blue is compatible with
mass spectrometry.
See Burgess Cassler, A. et. al. Clinical Chem.
35: 2297 1989 (pdf
download) for validation of our standard format gels.
PRICE 2D-ES-1: 2D ELECTROPHORESIS
standard format.
Our standard format gel
size is perfect
for immunoprecipitations, subcellular fractionations, most bacterial
preparations and many proteomic applications. The maximum load is 200 ug
protein in a maximum volume of 50 ul. One Coomassie blue stained, 2-D gel of dried
dimensions 16 cm wide x 13 cm long, is returned for each sample along with a pH
gradient plot and a method description sheet suitable for reports or
publications. The example below is from an Escherichia
coli sample prepared in our Urea sample buffer (left) and SDS Boiling
Buffer (right) run identically on a pH 4-8 IEF gradient and 10% acrylamide slab
gel. The great advantage of carrier ampholine (CA) 2D electrophoresis is
compatibility with SDS buffer and ease of sample preparation. Molecular weight standards (220,000, 94,000, 60,000, 43,000,
29,000, and 14,000) appear as bands on the right side of the gel labeled with
press-on numbers. One isoelectric point marker, added to each sample as an
internal standard, is marked with an arrow. Our standard IEF tube gel contains
2% pH 4-8 BDH ampholines; transparency drying (STP-3) is standard. The duplicate
gel rate is used for duplicate gels with different stains and also for blots,
but not for different gel conditions. See 2D-ES-5 below for our large format option.
Price: 2D-ES-1:
1 sample, $185; 2D-ES-1A:
2-3 samples, $175 each;
2D-ES-1B: 4-8 samples,
$165 each; 2D-ES-1C:
9-20 samples, $155 each;
2D-ES-1D: 21-200
samples, $147 each; 2D-duplicate: $90 each. More than 200 samples
- Inquire.
PRICE 2D-ES-5 LARGE FORMAT 2D ELECTROPHORESIS
The larger size gives better resolution for
proteomics applications; heavier loads may be applied to obtain more
material for mass spectrometry. The maximum load is 600 ug protein in a
maximum volume of 150 ul. One Coomassie blue-stained, dried 2D gel
of dimensions 20 cm wide by 22 cm long is returned for each sample along
with a pH gradient plot and a method description sheet. Our
standard IEF tube gel contains 2% pH 4-8 ampholines. Molecular
weight standards (220, 94, 60, 43, 29, and 14 kDa) appear as bands on
the right side of the gel labeled with press-on numbers. One IEF marker,
added to each sample as an internal standard, is marked with an arrow.
Transparency drying (STP-5, page 13) is standard. See above for Custom
pH gradients. Typical protein loads are 100 (silver staining) or 600 ug
(Coomassie blue) in 25-150 ul. LF Gel Price:
2D-ES-5: 1 sample $260; 2D-ES-5A: 2-3
samples $245 each; 2D-ES-5B:
4-12 samples $235 each; 2D-ES-5C:
13-200 samples $225 each, LF-Dup:
Large Format 2D duplicate gels: $105 each. More than 200 samples -
Inquire.
Call 800-462-3417 or email
2d@kendricklabs.com for a price
quote without obligation.
