Introduction
Bio-Rad’s
2-D SDS-PAGE standards are
Introduction
formulated
to provide
a two-dimensional
Bio-Rad’s 2-D
SDS-PAGE
standards are protein
pattern
with
detection
either
by silver or protein
formulated to provide a two-dimensional
Coomassie
pattern withstaining.
detection either by silver or
Coomassie
The
standardstaining.
proteins are defined by isolectric
point
(pl)
and
molecular
weight.
Added
to the
The standard proteins are
defined
by isolectric
sample,
used toweight.
determine
pls to
and
point (pl)they
and are
molecular
Added
the
molecular
weights
of sample
proteinspls
orand
to serve
sample, they
are used
to determine
as
a marker
for 2-Dofgel
matching.
Theorcharactermolecular
weights
sample
proteins
to serve
istic
pattern
of
main
and
minor
spots
each
as a marker for 2-D gel matching. Theofcharacterstandard
protein
makes
to of
identify
istic pattern
of main
andthem
minoreasy
spots
each
among
sample
standard
proteinspots.
makes them easy to identify
among
sample
spots.
In
addition,
the standards
can be used alone as
an
internal
control
for
reproducibility
of 2-D
In addition, the standards can be used
alone as
experiments
or to test
ampholyte mixtures.
an internal control
for reproducibility
of 2-D
experiments
or to advantage
test ampholyte
The
most striking
of themixtures.
standards is
that
valid
comparison
of 2-D
The they
mostallow
striking
advantage
of the
standards is
electrophoretic
patterns
betweenof
different
that they allow valid
comparison
2-D
laboratories.
electrophoretic patterns between different
2-D SDS-PAGE Standards,
Specifications
2-D SDS-PAGE Standards,
pl
range
4.5 to 8.5
Specifications
Molecular weight
pl range
range
Molecular weight
Contents
range
Contents
Volume
Shipping
Volume
Storage
Shipping
Storage
Shelf life
Applications per vial
Shelf life
Applications per vial
4.5 to 8.5
17,500
to 76,000
Hen egg white conalbumin type 1
17,500 serum
to 76,000
Bovine
albumin (BSA)
Hen egg
white actin
conalbumin type 1
Bovine
muscle
Bovine
serum
(BSA) 3-phosphate
Rabbit muscle albumin
glyceraldehyde
Bovine muscle actin
dehydrogenase
(GAPDH)
Rabbit carbonic
muscle glyceraldehyde
3-phosphate
Bovine
anhydrase
dehydrogenase
Soybean
trypsin (GAPDH)
inhibitor
Bovine
carbonic
anhydrase
Equine myoglobin
9Soybean
M urea trypsin inhibitor
Equine
myoglobin
5%
2-mercaptoethanol
9 MBio-Lyte
urea ® 5/7 ampholyte
2%
5%
2-mercaptoethanol
Distilled water
® 5/7
2% Bio-Lyte
ampholyte
500
µl solution,
ready
to use
Distilled water
4°C
500 µl solution, ready to use
-20°C
4°C
Aliquoting
is recommended prior to freezing. Frequent
-20°C
freeze/thaw
procedures should be avoided. Aliquots
Aliquoting
is recommended
prior
to week
freezing. Frequent
can
be stored
at 4°C for about
one
freeze/thaw procedures
shouldthe
be 2-D
avoided.
Aliquots
Important!
Always vortex
SDS-PAGE
can be stored atstandards
4°C for about
one
weekor
after thawing
Important!
Always vortex
2-D SDS-PAGE
refrigerating
to the
resuspend
the urea
standards after thawing or
One year
refrigerating
resuspendstain
the urea
200 (2.5 µl per mini
gel) usingtoCoomassie
One
year
1,000 (0.5–2.5 µl per mini gel) using silver stain
200 (2.5these
µl peramounts
mini gel)for
using
Coomassie
stain cm)
Double
full-length
gels (16–20
1,000 (0.5–2.5 µl per mini gel) using silver stain
Double these amounts for full-length gels (16–20 cm)
laboratories.
Bio-Rad Laboratories, Inc.
2000 Alfred Nobel Dr., Hercules, CA 94547 USA
Bio-Rad Laboratories, Inc.
(510) 741-1000
4110064 Rev A
2000 Alfred Nobel Dr., Hercules, CA 94547 USA
(510) 741-1000
4110064 Rev A
Instruction Manual
Instruction Manual
2-D SDS-PAGE
2-D SDS-PAGE
Standards
Standards
Bio-Rad Laboratories, Inc.
2000 Alfred Nobel Dr., Hercules, CA 94547 USA
Bio-Rad Laboratories, Inc.
(510) 741-1000
4110064 Rev A
2000 Alfred Nobel Dr., Hercules, CA 94547 USA
(510) 741-1000
4110064 Rev A
Instruction Manual
Instruction Manual
2-D SDS-PAGE
2-D SDS-PAGE
Standards
Standards
Ordering Information
Catalog # Description
Ordering Information
161-0320 2-D SDS-PAGE Standards
Catalog # Description
161-0303 SDS-PAGE Standards, high range
161-0320 2-D SDS-PAGE Standards
161-0304 SDS-PAGE Standards, low range
161-0303 SDS-PAGE Standards, high range
161-0363 Precision Plus Protein™ Unstained Standards
161-0304 SDS-PAGE Standards, low range
161-0373 Precision Plus Protein ™
All Blue Standards
161-0363 Precision Plus Protein Unstained Standards
161-0374 Precision Plus Protein Dual Color Standards
161-0373 Precision Plus Protein All Blue Standards
161-0374 Precision Plus Protein Dual Color Standards
Catalog # Description
Ordering Information
161-0320 2-D SDS-PAGE Standards
Catalog # Description
161-0303 SDS-PAGE Standards, high range
161-0320 2-D SDS-PAGE Standards
161-0304 SDS-PAGE Standards, low range
161-0303 SDS-PAGE Standards, high range
161-0363 Precision Plus Protein™ Unstained Standards
161-0304 SDS-PAGE Standards, low range
161-0373 Precision Plus Protein All Blue Standards
161-0363 Precision Plus Protein™ Unstained Standards
161-0374 Precision Plus Protein Dual Color Standards
161-0373 Precision Plus Protein All Blue Standards
161-0374 Precision Plus Protein Dual Color Standards
Ordering Information
Two-Dimensional Electrophoretic Protein Pattern
Molecular weights
8.5
7.0
5.9
6.6
5.5
5.1
4.5
pl values of standard proteins
Protein spots
1 Conalbumin
76,000
66,200
2
43,000
2 BSA
1
3 Actin
3
36,000
31,000
4
5
17,500
4 GAPDH
5 Carbonic anhydrase
21,500
6
7
6 Trypsin inhibitor
7 Myoglobin
Two-dimensional electrophoretic protein pattern
of 2-D SDS-PAGE standards separated in the

Mini-PROTEAN II cell. 2.5 µl of the standards
were run according to the method of Klose
(Klose J, Humangenetik 26, 231–243, 1975;
Klose K and Feller M, Electrophoresis 2, 12–24,
1981; Jungblut PR and Seifert R, J Biochem
Biophys Methods 21, 47–58, 1990)
Protein Molecular Weights and
Isoelectric Points (pl)
Protein
Molecular
Weight
pl
Reference
Protein
Molecular
Weight
pl
Reference
Hen egg white
conalbumin
type 1
76,000
6.0
6.3
6.6
Cavorta P, et al., Experientia 34,
849–850 (1978)
Bovine carbonic
anhydrase
31,000
5.9
6.0
Bovine serum
albumin (BSA)
66,200
5.4
5.6
Brown JR, Fed Proc 34, 591 (1975)
Bovine muscle
actin
43,000
5.0
5.1
The running pi value of muscle actin has
been empirically determined.
Ashworth RB, et al., Arch Biochem
Biophys 142, 122–131 (1971)
Jonsson M and Petterson E, Acta Chem
Scand, 22, 712–713 (1986)
Davis RP, Carbonic Anhydrase,
p. 545–564 in The Enzymes, Vol V, 2nd
edn (Boyer PD, ed), Academic Press,
New York (1971)
4.5
36,000
8.3
8.5
Smith CM and Velick SF, J Biol Chem 247,
273–284, (1972)
Soybean trypsin
Inhibitor
21,500
Rabbit muscle
GAPDH
Catsimpoulas N and Leuthner E, Anal
Biochem 31, 437–447 (1969)
Wu YV and Scheraga HG, Biochemistry 1,
698–705 (1962)
Equine
myoglobin
17,500
7.0
Salaman MR and Williamson AR,
Biochem J 122, 93–99 (1971)