Designed for optimal performance
and confidence in your results
Novex® protein separation, blotting, and detection systems
Thoughtfully designed products for
accurate, reproducible results
Novex® protein separation, blotting,
and detection systems
ǩSuperior protein resolution from advanced
separation and transfer technologies
ǩUltrasensitive protein detection
ǩRapid, consistent results from run to run
Today the Novex® brand includes more
than just gels. It also covers high-quality
products and reagents designed to work
with the original gel product line, including
the sharpest, most accurate prestained
and unstained molecular weight standards;
uniquely formulated, sensitive stains;
the iBlot® 7-Minute Blotting System, the
BenchPro® 4100 Western Processing System,
and western detection kits. Using these
products together helps ensure complete
protein separation with tight band resolution,
accurate molecular weight estimation,
complete transfer, easy post-transfer
processing, and sensitive detection.
2
Contents
Protein separation
4 Electrophoresis and precast gels
9 Gel casting essentials
10 Large protein separation
11 Standards
13 Stains
Western transfer
15 Transfer equipment
Western detection
17 Detection equipment
18 Detection essentials
3
Gel electrophoresis
High-performance, high-quality precast gels and gel casting
essentials
If you are doing standard one-dimensional protein separation,
we have Novex® solutions, and this guide will help you select
the separation tools that fit your research. The Novex® gel
line consists of several different chemistries, well formats,
and gel sizes, so you get the separation you need for accurate
downstream results.
One-dimensional protein separation.
Gel casting accessories
Tris-glycine gels
NuPAGE® gels
Application
Pour-your-own essentials:
preassembled empty cassettes, buffers,
reagents
Precast convenience with
Laemmli chemistry
Straight protein bands,
longest shelf life
Protein resolution
Medium
Medium
High
Protein integrity
Medium
Medium
High
Shelf life
NA
1–2 months
8 months (Tris-acetate)
16 months (Bis-Tris)
Average run time
90 min
90 min
35 min
Separation range
6–500 kDa
6–500 kDa
1.5–300 kDa (Bis-Tris)
30–400 kDa (Tris-acetate)
Gel % available
NA
4%, 6%, 8%, 10%, 12%, 4–20%, 4–12%,
14%, 16%, 18%, 8–16%, 10–20%
4–12%, 8%, 10%, 12% (Bis-Tris)
3–8%, 7% (Tris-acetate)
Pack sizes available
NA
5- and 10-packs
2- and 10-packs
Gel dimensions
Mini: 8 x 8 cm, 1 or 1.5 mm thick
Midi: 8 x 13 cm, 1 mm thick
Mini: 8 x 8 cm, 1 or 1.5 mm thick
Midi: 8 x 13 cm, 1 mm thick
Mini: 8 x 8 cm, 1 or 1.5 mm thick
Midi: 8 x 13 cm, 1 mm thick
Maximum sample
volume/well
25 µL (1 mm)
37 µL (1.5 mm)
25 µL (1 mm)
37 µL (1.5 mm)
25 µL (1 mm)
37 µL (1.5 mm)
Maximum protein
load per band for
optimal resolution*
NA
0.5 µg
0.5 µg
* On 1.0 mm, 10-well gel, detected by Coomassie staining.
4
®
NuPAGE Gels
The NuPAGE® precast gel system is a revolutionary highperformance polyacrylamide gel system.
Gels are available in both Bis-Tris and Tris-acetate
formulations and in a variety of acrylamide percentages
(Table 1, Figure 2). The unique NuPAGE® gel formulations
minimize the “smiles” and poor resolution seen with other
gel types. The neutral-pH gel chemistry and buffer system
avoid chemical sample modifications that occur in alkaline
environments. In addition, proteins transfer from NuPAGE®
gels more efficiently, resulting in higher detection signals
during western analysis.
ǩ Superior protein band resolution and stability
ǩ Faster sample run times
ǩ Longer product shelf life
Table 1. Two NuPAGE® types to choose from.
Gel type
Percentages availablex
Separation range
Shelf life
Average run time
Applications
Bis-Tris
8%, 10%, 12%, 4–12%
1.5–300 kDa
16 months*
35–50 min
Separation of small- to medium-sized
proteins; simple sample reduction; protein
sequencing, mass spectrometry, techniques
where protein integrity is crucial
Tris-acetate
7%, 3–8%
30–400 kDa
8 months *
60 min
Excellent separation of large molecular
weight proteins
Ordering information for NuPAGE® Mini Gels. Each box of NuPAGE® Mini Gels includes 10 gels. For additional pack sizes, please visit our website
at www.lifetechnologies.com/nupage
1.0 mm thickness
1.5 mm thickness
Gel type and percentage
10 wells
12 wells
15 wells
2D well
10 wells
15 wells
Bis-Tris 10%
NP0301BOX
NP0302BOX
NP0303BOX
NP0306BOX
NP0315BOX
NP0316BOX
NP0317BOX
Bis-Tris 4–12%
NP0321BOX
NP0322BOX
NP0323BOX
NP0326BOX
NP0335BOX
NP0336BOX
NP0337BOX
Bis-Tris 12%
NP0341BOX
NP0342BOX
NP0343BOX
NP0346BOX
Tris-acetate 7%
EA0355BOX
EA03552BOX EA03555BOX
EA0358BOX
EA03585BOX EA0359BOX
Tris-acetate 3–8%
EA0375BOX
EA3752BOX
EA0378BOX
EA03785BOX EA0379BOX
EA03755BOX EA0376BOX
2D well
Ordering information for NuPAGE® Midi Gels. Each box of NuPAGE® Midi Gels includes 10 gels.
Without Midi Gel Adapters
With Midi Gel Adapters (10)*
Gel type and percentage
12+2 wells
20 wells
26 wells
12+2 wells
20 wells
26 wells
Bis-Tris 8%
WG1001BOX
WG1002BOX
WG1003BOX
WG1001A
WG1002A
WG1003A
Bis-Tris 10%
WG1201BOX
WG1202BOX
WG1203BOX
WG1201A
WG1202A
WG1203A
Bis-Tris 4–12%
WG1401BOX
WG1402BOX
WG1403BOX
WG1401A
WG1402A
WG1403A
Tris-acetate 3–8%
WG1601BOX
WG1602BOX
WG1603BOX
WG1601A
WG1602A
WG1603A
* By attaching the Midi Gel Adapter (Cat. No. WA0999) to the gel cassette, you can run the midi gels in the Bio-Rad Criterion™ Cell.
5
A
B
Gel tanks and buffers
All Novex® gels are available in the 8 x 8 cm mini gel format
and a larger 8 x 13 cm midi size for higher throughput. Run
up to two mini gels at a time in the XCell SureLock® Mini-Cell
(Figure 1A). Use the XCell4 SureLock™ Midi-Cell (Figure 1B)
to simultaneously run up to four midi gels. Each midi gel can
run up to 26 samples, allowing electrophoresis of over 100
samples at a time. With a Midi Gel Adapter (Cat. No. WA0999),
Novex® Midi Gels can be run in a Bio-Rad Criterion™ Cell.
Figure 1. The Xcell SureLock® protein electrophoresis cells. (A) XCell
SureLock® Mini-Cell (Cat. No. EI0001) for running one or two NuPAGE®
or Novex® mini gels. (B) XCell4 SureLock™ Midi-Cell (Cat. No. WR0100)
for running up to four NuPAGE® or Novex® midi gels.
Product
Quantity
Cat. No.
1
EI0001
XCell4 SureLock Midi-Cell
1
WR0100
PowerEase® 500 Power Supply
1
EI8600
250 µL
NP0004
10 mL
NP0009
NuPAGE Antioxidant
15 mL
NP0005
NuPAGE® LDS Sample Buffer (4X)
10 mL
NP0007
250 mL
NP0008
500 mL
NP0001
5L
NP0001-02
500 mL
NP0002
5L
NP0002-02
500 mL
LA0041
NuPAGE MOPS SDS Buffer Kit (for Bis-Tris gels) contains 1 each of NP0001, NP0004, NP0005, NP0007
1 kit
NP0050
NuPAGE® MES SDS Buffer Kit (for Bis-Tris gels) contains 1 each of NP0002, NP0004, NP0005, NP0007
1 kit
NP0060
NuPAGE Tris-Acetate SDS Buffer Kit (Tris-acetate gels only) contains 1 each of LA0041, NP0004, NP0005, NP0007
1 kit
LA0050
NuPAGE® Transfer Buffer (20X)
125 mL
NP0006
1L
NP0006-1
®
XCell SureLock Mini-Cell
™
®
NuPAGE Sample Reducing Agent (10X)
®
NuPAGE® MOPS SDS Running Buffer (for Bis-Tris gels only) (20X)
NuPAGE® MES SDS Running Buffer (for Bis-Tris gels only) (20X)
NuPAGE® Tris-Acetate SDS Running Buffer (20X)
®
®
Upgrade your gels to NuPAGE® gels
If you would like to upgrade to NuPAGE® gels from Tris-glycine gels, visit us at lifetechnologies.com/nupage for an easy conversion
table.
6
6
Tris-acetate†
Bis-Tris*
8%
Bis-Tris Gel
w/MES
running
buffer
8%
Bis-Tris Gel
w/ MOPS
running
buffer
10%
Bis-Tris Gel
w/ MES
running
buffer
10%
Bis-Tris Gel
w/ MOPS
running
buffer
4–12%
Bis-Tris Gel
w/ MES
running
buffer
4–12%
Bis-Tris Gel
w/ MOPS
running
buffer
12%
Bis-Tris Gel
w/ MES
running
buffer
12%
Bis-Tris Gel
w/ MOPS
running
buffer
3–8%
Tris-acetate
Gel w/ TA
running
buffer
7%
Tris-acetate
Gel w/ TA
running
buffer
0%
260 kDa
260 kDa
160 kDa
260 kDa
260 kDa
10%
260 kDa
260 kDa
160 kDa
260 kDa
20%
160 kDa
110 kDa
80 kDa
160 kDa
110 kDa
60 kDa
30%
110 kDa
160 kDa
50 kDa
80 kDa
40 kDa
60 kDa
60 kDa
110 kDa
160 kDa
50 kDa
160 kDa
80 kDa
110 kDa
60 kDa
80 kDa
50 kDa
500 kDa
500 kDa
60 kDa
290 kDa
50 kDa
110 kDa
290 kDa
240 kDa
160 kDa
240 kDa
116 kDa
30 kDa
40 kDa
97 kDa
80 kDa
20 kDa
50 kDa
160 kDa
40 kDa
30 kDa
50 kDa
110 kDa
40 kDa
80 kDa
60 kDa
80 kDa
40%
160 kDa
110 kDa
260 kDa
80 kDa
60 kDa
50%
40 kDa
20 kDa
40 kDa
30 kDa
30 kDa
50 kDa
60 kDa
60%
30 kDa
15 kDa
15 kDa
20 kDa
55 kDa
40 kDa
20 kDa
66 kDa
15 kDa
20 kDa
15 kDa
66 kDa
97 kDa
10 kDa
50 kDa
70%
116 kDa
10 kDa
30 kDa
30 kDa
3.5 kDa
10 kDa
40 kDa
55 kDa
15 kDa
40 kDa
80%
10 kDa
20 kDa
20 kDa
30 kDa
90%
3.5 kDa
3.5 kDa
15 kDa
3.5 kDa
10 kDa
20 kDa
40 kDa
15 kDa
10 kDa
10 kDa
100%
* Migration patterns of Novex® Sharp Protein Standards (Cat. No. LC5800, Pre-stained; Cat. No. LC5801, Unstained) on NuPAGE® Bis-Tris gels.
†
Migration patterns of HiMark™ Unstained Standard (Cat. No. LC5688) on NuPAGE® Tris-acetate gels.
Figure 2. Migration patterns achieved in NuPAGE® gels. For optimal results, protein bands should migrate within the gray shaded areas.
To learn more about additional Novex® gel formulations, percentages, and formats, go to lifetechnologies.com/novex1d
7
Precast Laemmli-style gels for protein analysis—
Tris-glycine gels
Novex® Tris-glycine polyacrylamide gel chemistry is based on the Laemmli system, with minor modifications for maximum
performance in the precast format. These gels do not contain SDS and can therefore be used to accurately separate both native
and denatured proteins. Tris-glycine gels provide reproducible separation of a wide range of proteins into well-resolved bands
(Figure 3).
Tris-Glycine Gels
†
Large proteins*
(116–500 kDa)
4%
0%
6%
Small proteins†
(3–60 kDa)
Mid-size proteins
(20–250 kDa)
8%
10%
12%
14%
16%
Wide range†
(6–200 kDa)
18%
4–12%
8–16%
4–20%
10–20%
500 kDa
260kDa
kDa
260
500 kDa
10%
260 kDa
260 kDa
290 kDa
20%
260 kDa
260 kDa
160 kDa
160 kDa
30%
240 kDa
160 kDa
260 kDa
110 kDa
290 kDa
110 kDa
80 kDa
60 kDa
160 kDa
110 kDa
80 kDa
50 kDa
40%
60 kDa
240 kDa
80 kDa
110 kDa
160 kDa
160 kDa
110 kDa
110 kDa
80 kDa
80 kDa
60 kDa
60 kDa
50 kDa
40 kDa
50 kDa
260 kDa
20 kDa
20 kDa
20 kDa
40 kDa
15 kDa
15 kDa
10 kDa
40 kDa
110 kDa
80 kDa
50 kDa
40 kDa
50 kDa
30 kDa
30 kDa
20 kDa
40 kDa
60 kDa
20 kDa
15 kDa
50 kDa
40 kDa
80%
60 kDa
60 kDa
10 kDa
30 kDa
50 kDa
80 kDa
110 kDa
60 kDa
110 kDa
160 kDa
110 kDa
50 kDa
97 kDa
60 kDa
40 kDa
60%
160 kDa
160 kDa
160 kDa
30 kDa
70%
80 kDa
30 kDa
30 kDa
110 kDa
260 kDa
40 kDa
60 kDa
80 kDa
116 kDa
260 kDa
40 kDa
50 kDa
160 kDa
50%
160 kDa
260 kDa
30 kDa
10 kDa
15 kDa
15 kDa
50 kDa
10 kDa
20 kDa
90%
166 kDa
10 kDa
66 kDa
30 kDa
40 kDa
100%
30 kDa
15 kDa
10 kDa
20 kDa
55 kDa
97 kDa
20 kDa
15 kDa
10 kDa
15 kDa
Figure 3. Migration patterns of protein molecular weight standards in Tris-glycine gels. For optimal results, protein bands should
migrate within the gray shaded areas. Tris-glycine gels are available in a variety of acrylamide percentages, and in the mini or midi format.
*Migration patterns of HiMark™ Unstained Standard (Cat. No. LC5688) on Tris-Glycine Gels.
†Migration patterns of Sharp Protein Standards (Cat. No. LC5800, Pre-stained; Cat. No. LC5801, Unstained) on Tris-Glycine Gels.
To learn more about Novex® gels, go to lifetechnologies.com/buyproteingels
8
®
Novex gel casting equipment
From high-purity acrylamide to gel combs, our do-it-yourself
ingredients and equipment make gel casting easy and efficient.
Product
Quantity
Cat. No.
200 mL
R-33400
30 mL
15524-010
UltraPure Acrylamide
500 g
15512-023
UltraPure™ N,N´-Methylenebisacrylamide
100 g
15516-024
™
1 kg
15504-020
™
UltraPure Sucrose
5 kg
15503-022
UltraPure™ 10% SDS Solution
4 x 100 mL
15553-027
1L
24730-020
500 g
15525-017
Cassettes, 1.0 mm
25/pack
NC2010
Cassettes, 1.5 mm
25/pack
NC2015
12 + 2 well
10/pack
WC1012
Combs, 1.0 mm, 1 well
25/pack
NC3001
Combs, 1.0 mm, 10 well
25/pack
NC3010
Combs, 1.0 mm, 12 well
25/pack
NC3012
Combs, 1.0 mm, 15 well
25/pack
NC3015
Combs, 1.0 mm, 2 well
25/pack
NC3003
Combs, 1.0 mm, 2D well
25/pack
NC3002
Combs, 1.0 mm, 5 well
25/pack
NC3005
Combs, 1.5 mm, 10 well
25/pack
NC3510
Combs, 1.5 mm, 15 well
25/pack
NC3515
Combs, 1.5 mm 2D well
25/pack
NC3502
Midi Cassette Combs, 20 well
10/pack
WC1020
Midi Cassette Combs, 26 well
10/pack
WC1026
500 g
15505-035
2 kg
15505-050
Acrylamide, polymerizing agents, and buffers
Rhinohide™ polyacrylamide gel strengthener concentrate,
sufficient additive for 1 L of 30% acrylamide/bis-acrylamide (37.5:1)
UltraPure™ TEMED
™
UltraPure Tris
™
UltraPure SDS
Gel cassettes
Gel combs
Denaturing agents
UltraPure™ Urea
Visit our website at www.lifetechnologies.com/novexessentials for more products.
9
Reliable analysis of very large protein complexes
Native Complex Purification and Analysis System
ǩ NativePAGE™ Bis-Tris gels for sensitive, high-resolution
separations
ǩ NativeMARK™ Unstained Protein Standard for accurate
molecular weight estimation
ǩ NativePure™ Purification Systems for efficient purification of
native protein complexes
The Native Complex Purification and Analysis System is a novel
set of tools for the analysis of very large protein complexes,
membrane protein complexes, and super complexes.
™
NativePAGE Bis-Tris Gel System
The NativePAGE™ Bis-Tris Gel System is a precast
polyacrylamide mini gel system that provides a sensitive and
high-resolution method for analysis of native membrane
protein complexes and native soluble proteins, molecular
mass estimations, and assessment of native protein purity. It
is based on the blue-native polyacrylamide gel electrophoresis
technique developed by Schägger and von Jagow. Use this
system to maximize resolution of large proteins (>10,000 kDa),
analyze membrane protein complexes in their native,
undenatured conformations, and obtain better resolution than
with traditional Tris-glycine native electrophoresis (Figure 4).
Product
NativeMARK™ Unstained
Protein Standard
The NativeMARK™ Unstained Protein Standard is a ready-touse protein marker that enables accurate molecular weight
estimation of proteins using NativePAGE™, Tris-glycine, or
NuPAGE® Tris-acetate gels. Proteins resolve into distinct bands
in the range of ~20–1,200 kDa.
A
kDa 1
1,236
1,048
720
480
242
146
B
2
3
4
5
kDa 1
2
3
4
5
1,236
1,048
720
480
242
66
146
66
20
20
Figure 4. Improved separation with NativePAGE™ gels. Native
electrophoresis was performed with (A) 4–12% Tris-glycine gels and
(B)NativePAGE ™ 4–16% gels. Both gels were loaded with NativeMARK ™
standards (lane 1) and 18 mg spinach chloroplast extract solubilized in
0.25%, 0.5%, 1.0%, and 2.0% dodecylmaltoside (lanes 2–5). Gels were
stained with the Colloidal Blue Staining Kit.
Quantity
Cat. No.
10 gels
BN1001
NativePAGE 3–12% Bis-Tris Gels (15 well)
10 gels
BN1002
NativePAGE™ 4–16% Bis-Tris Gels (10 well)
10 gels
BN1003
Native protein analysis
NativePAGE™ 3–12% Bis-Tris Gels (10 well)
™
™
NativePAGE 4–16% Bis-Tris Gels (15 well)
10 gels
BN1004
NativeMARK™ Unstained Protein Standard
250 µL
LC0725
NativePAGE™ Running Buffer Kit
1 kit
BN2007
NativePAGE Running Buffer Kit
1 kit
BN2008
NativePAGE™ 20X Running Buffer
1L
BN2001
™
250 mL
BN2002
™
10 mL
BN2003
™
0.5 mL
BN2004
™
NativePAGE 20X Cathode Buffer Additive
NativePAGE 4X Sample Buffer
NativePAGE 5% G-250 Sample Additive
* NativePAGE™ gels are available in an 8 x 8 cm mini gel format.
NativePure™ Affinity Purification Kits
NativePure™ Affinity Purification Kits allow you to easily purify interacting protein complexes for analysis. There are two kits
currently available: the NativePure™ Mammalian Affinity Purification Kit and the NativePure™ Lentiviral Expression and Affinity
Purification System. Both offer Gateway® vectors for simplified expression in multiple expression systems. To learn more about
native protein analysis, go to lifetechnologies.com/native
10
Accurate molecular weight estimations
The sharpest, most accurate prestained and unstained MW standards
Sharp prestained and unstained protein standards
Protein separation is often accompanied by band identification.
A variety of Novex® molecular weight (MW) protein standards
are available to help you make the most accurate size
estimations possible.
Our complete line of Novex® protein molecular weight markers
is designed for convenience and accuracy in electrophoresis
band identification.
To learn more about protein molecular weight standards,
go to lifetechnologies.com/proteinstandards
ǩ Sharpest bands—clearer estimation of MW compared to
thick bands
ǩ Broadest MW range—estimation over a larger range
(3.5–260 kDa)
ǩ Ladder-like separation of bands—evenly spaced pattern
across the readable range for easy identification and
comparison
SeeBlue® Pre-stained
Standard
BenchMark™ Prestained Protein
Ladder
HiMark™ Pre-stained
Protein Standard
SeeBlue® Plus2 Prestained Standard
Sharp Pre-stained
Protein Standard
Application
Basic, routine MW
MW marker for Trisglycine gels only
Ideal for large proteins
Multiple color bands,
easy estimation
Easiest to interpret
MW; best size
estimation
# of bands/
9 bands
10 bands
9 bands
10 bands
12 bands
# of colors
1 color
2 colors
2 colors
3 colors
12 colors
Band sharpness
Low
Low
Low
Medium
High
MW range
4–250 kDa
10–190 kDa
30–460 kDa
4–250 kDa
Gel compatibility
®
NuPAGE Bis-Tris
Tris-glycine only
Tricine
3.5–260 kDa
®
Tris-acetate
NuPAGE Bis-Tris
NuPAGE® Bis-Tris
Tris-glycine
Tricine
Tricine
Tris-acetate
Tris-glycine
Tris-glycine
Tris-glycine
Pre-stained protein standards
for easy and clear band identification
kDa
Product
Quantity
kDa
Cat. No.
kDa
kDa
kDa
188
460
198
260
Pre-stained protein standards
SeeBlue®
98
160
500 µL
110
80
LC5625
2 x 250 µL
62
171
~85
60
49
~60
49
38
~40
28
30
117
~50
38
40
10748-010
~120
62
50
BenchMark™
268
238
~190
28
HiMark™
250 µL
20
LC5699
~25
18
17
15
71
~20
14
14
SeeBlue® Plus2
500 µL
LC5925
10
2 x 250 µL
55
~10
41
6
6
3.5
Sharp
~15
LC5800
3
3
Standard
Cat. No.
Sharp
Pre-stained
31
SeeBlue®
Plus2
SeeBlue®
BenchMark™
Pre-stained
HiMark™
Pre-stained
LC5800
LC5925
LC5625
10748-010
LC5699
NuPAGE®
4–12% Bis-Tris
Gel w/ MES
SDS buffer
NuPAGE®
4–12% Bis-Tris
Gel w/ MES
SDS buffer
NuPAGE®
4–12% Bis-Tris
Gel w/ MES
SDS buffer
4–20%
Tris Glycine Gel
NuPAGE® 3–8%
Tris-Acetate Gel
w/ Tris-acetate
SDS buffer
11
Unstained protein standards
NativeMark™
Unstained Protein
Standard
Mark12™ Unstained
Standard
BenchMark™ Protein
Ladder
Sharp Unstained Protein
Standard
HiMark™ Unstained
Protein Standard
Application
Routine MW estimation in denaturing gel
electrophoresis
MW estimation and quick
orientation
MW estimation and band
identification
MW estimation for
large proteins
MW estimation
in native gel
electrophoresis
Number of bands
12
15
12
9
8
MW range
2.5–200 kDa
10–220 kDa
3.5–260 kDa
40–500 kDa
20–1,200 kDa
Orientation band(s)
No
Yes
Yes
No
No
Number of orientation band(s)
0
2 (20 and 50 kDa bands
more prominent)
3 (20, 50, and 110 kDa
bands more prominent)
0
0
Gel compatibility
NuPAGE® Bis-Tris;
SDS-PAGE Tricine, Trisacetate, Tris-glycine
NuPAGE® Bis-Tris;
SDS-PAGE Tricine, Trisacetate, Tris-glycine
NuPAGE® Bis-Tris;
SDS-PAGE Tricine, Trisacetate, Tris-glycine
Tris-acetate
Tris-glycine,
NativePAGE™ BisTris, Tris-acetate
Band sharpness
Low
Medium
High
Low
Low
Unstained protein standards
Quantity
Cat. No.
Sharp
2 x 250 µL
LC5801
2 x 250 µL
10747-012
1 mL
LC5677
HiMark HMW
250 µL
LC5688
MagicMark™ XP Western
250 µL
LC5602
BenchMark™ His-tagged
125 µL
LC5606
BenchMark™ Fluorescent
125 µL
LC5928
IEF Marker 3–10
500 µL
39212-01
250 µL
LC0725
™
BenchMark Ladder
Mark12™
™
™
NativeMark
Unstained protein standards for
accurate molecular weight estimation
kDa
kDa
kDa
260
220
200
160
160
110
80
120
100
90
80
70
60
Protein standard
for western blots
kDa
Specialty
protein standards
kDa
kDa
IEF marker
kDa
kDa
pI
8.3
8.0
500
60
50
40
30
15
25
120
100
116
97
50
50
7.4
40
32
6.9
21
6.0
146
20
20
66
5.3
5.2
11
4.5
15
55
242
30
15
3.5
2.5
10
480
60
40
30
66
15
63
30
40
6
3.5
720
98
20
20
kDa
1,048
7.8
155
40
14.4
10
120
80
50
31
21.5
160
120
60
60
160
160
80
80
36.5
40
20
290
240
116.3
97.4
66.3
55.4
50
30
220
Native
marker
10
4.2
20
10
40
3.5
Standard
Novex® Sharp
BenchMark™
Mark12™
HiMark™
MagicMark™ XP
Cat. No.
LC5801
10747-012
LC5677
LC5688
LC5602
NuPAGE® 4–12%
Bis-Tris Gel w/ MES,
stained with
Coomassie® R-250
NuPAGE® 3–8%
Tris-Acetate Gel
w/ Tris-acetate
SDS buffer stained
with SimplyBlue™
SafeStain
NuPAGE® Bis-Tris Gel,
blotted to
nitrocellulose,
detected w/
WesternBreeze®
Chemiluminescent Kit
NuPAGE® 4–12%
NuPAGE® 4–12%
Bis-Tris Gel w/ MES, Bis-Tris Gel w/ MES,
stained with
stained with
SimplyBlue™ SafeStain Coomassie® R-250
12
Standard
Cat. No.
BenchMark™
His-tagged
LC5606
NuPAGE® 4–12%
NuPAGE® 4–12%
Bis-Tris Gel w/ MES,
Bis-Tris Gel w/ MES
stained w/
SDS buffer, blotted to
InVision™
nitrocellulose, detected
In-Gel His-tag
w/ anti-His (C-term)
Stain
antibody and
WesternBreeze®
Chemiluminescent Kit
BenchMark™
Fluorescent
IEF Marker
3–10
NativeMark™
Unstained
LC5928
39212-01
LC0725
NuPAGE® 4–12%
Bis-Tris Gel w/ MES,
visualized with
UV transilluminator
(2 sec exposure)
pH 3–10 IEF Gel
NativePAGE™
4–16%
Bis-Tris Gel
Sensitive stains for total detection
Life Technologies offers a number of staining reagents for total protein and posttranslationally modified proteins, including
Coomassie™ G-250 stain, silver stain, and a suite of compatible and highly sensitive fluorescent stains that allow direct
quantitation of differential phosphorylation and glycosylation patterns as well as total-protein expression from a single
sample on the same gel. Use the table below to find the stain that’s right for your application.
An overview of Novex® protein stains.
Coomassie Fluor™
Orange Protein Gel
Stain
SimplyBlue™ SafeStain
SilverQuest™ Silver Staining Kit
SYPRO® Ruby Protein Gel Stain
Type of stain
Colorimetric; ready-to-use
colloidal Coomassie™ G-250
Colorimetric; ready-to-use
silver
Fluorescent; ready-to-use
Fluorescent;
ready-to-use
Instrumentation
required
No
No
UV transilluminator, blue-light
box, or laser scanner
UV transilluminator,
blue-light box, or laser
scanner
Sensitivity
>7 ng BSA
Subnanogram
Subnanogram
4–8 ng
Background
Low
Low
Low
Low
Linear dynamic
range
7–70 ng
1–10 ng
0.25–1,000 ng
4–1,000 ng
Multiplex capability
No
No
Yes
No
For densitometry
Yes
Not recommended
Integration of fluorescent signal
Integration of fluorescent signal
MS compatibility
Yes
Yes
Yes
Yes
Staining
membranes
PVDF
No
Use SYPRO® Ruby Protein
Blot Stain
No
Total staining/
destaining time
3 hr; 12 min with microwave
protocol
90 min; <60 min with microwave protocol
90 min (mini gels with new
protocol); overnight for largeformat gels
30–60 min
Protocol summary
Rinse, stain, water-based
destain
10 solution changes
Fix, stain, and wash
Fix and stain in one step
Cat. No.
LC6060, LC6065
LC6070
S12001, S12000, S21900
C33250, C33251
Ultrasensitive fluorescent detection
of total protein
SYPRO® Ruby Protein Gel Stain
ǩ Simple staining procedure
ǩ Linear quantitation range of 3 orders of magnitude
ǩ Linear compatibility with mass spectrometry and
microsequencing
SYPRO® Ruby Protein Gel Stain is a highly sensitive fluorescent
stain for proteins in 1D or 2D gels (Figure 5). The fast
microwave mini gel protocol yields optimal results, typically
in only 90 minutes. For convenience, SYPRO® Ruby stain is
provided ready to use and stores at room temperature.
Figure 5. Sensitive fluorescent staining with SYPRO®
Ruby Protein Gel Stain.
13
Fast, sensitive, and safe
Coomassie™ staining with
SimplyBlue™ SafeStain
SimplyBlue™ SafeStain is a uniquely formulated, readyto-use Coomassie™ G-250 stain specifically designed for
fast, sensitive, and safe protein staining. With SimplyBlue™
SafeStain, bands develop in minutes using the rapid microwave
protocol, and as little as 7 ng of protein can be detected
(Figure 6). SimplyBlue™ SafeStain is completely nonhazardous
and does not require acid/alcohol fixative or destaining.
SimplyBlue™ SafeStain is compatible with mass spectrometry.
Figure 6. Sensitive staining with
SimplyBlue™ SafeStain.
Mass spectrometry–compatible
SilverQuest™ Silver Staining Kit
The SilverQuest™ Silver Staining Kit provides silver staining
sensitivity with minimal protein modification for optimal mass
spectrometry results. SilverQuest™ stain allows detection of
as little as 0.3 ng of protein (Figure 7), typically in less than 90
minutes. The optimized microwave procedure yields results in
less than 30 minutes.
Product
Figure 7. Subnanogram sensitivity with the
SilverQuest™ Silver Staining Kit.
Quantity
Cat. No.
1L
LC6060
3.5 L
LC6065
1 kit
LC6070
200 mL
S12001
1L
S12000
5L
S21900
1L
C33250
5L
C33251
1 kit
P33300
1 kit
P21857
Protein stains
SimplyBlue™ SafeStain
™
SilverQuest Silver Staining Kit
®
SYPRO Ruby Protein Gel Stain
™
Coomassie Fluor Orange Protein Gel Stain
Pro-Q® Diamond Phosphoprotein Gel Staining Kit
®
Pro-Q Emerald 300 Glycoprotein Staining Kit
14
Efficient protein transfer for successful
western blotting
A
®
iBlot 7-Minute Blotting System
1
B
2
3
4 5
6 7
8 9 10 11 12
1 2 3 4
5
6
7
8 9 10 11 12
ǩ Complete transfer in 7 minutes or less
ǩ High blotting efficiency and evenness
ǩ Increased blotting reliability and reproducibility
The iBlot® 7-Minute Blotting System is designed to efficiently
and reliably blots proteins from polyacrylamide gels in 7
minutes or less, without the need for buffers or an external
power supply. A self-contained unit (Figure 9), the iBlot® device
uses disposable blotting stacks with integrated nitrocellulose
or PVDF transfer membranes, offering the convenience
of a bufferless, plug-and-play system. In addition to ease
and convenience, the iBlot® system offers high-efficiency
protein transfers (Figure 8) for high downstream detection
sensitivities—in many cases, higher than conventional
methods. In the end, you can achieve more accurate detection
using less sample.
Figure 8. High transfer efficiencies achieved using the iBlot®
7-Minute Blotting System. (A) iBlot® dry transfer to nitrocellulose and
(B) semi-dry transfer to nitrocellulose of NuPAGE® 12% Bis-Tris Mini
Gels. Lanes 1–6: 0.0625 µg, 0.125 µg, 0.25 µg, 0.5 µg, 1.0 µg, and 2.0 µg
of SW480 human colon cancer cell lysate (Alexis) (control); lanes 7
and 12: 5 µL SeeBlue® Plus2 Pre-stained Protein Standard; lanes
8–11: 0.5 µL, 1.0 µL, 2.0 µL, and 4.0 µL of MagicMark™ XP Western
Protein Standard.
The iBlot® 7-Minute Blotting System is designed to work with
multiple polyacrylamide gel sizes, including the midi size,
fitting midi gels (8 x 13 cm) and E-PAGE™ 48 and 96 gels, and
mini gels (8 x 8 cm).
Precut blotting membranes
Simplify blotting setup with precut membranes/filter-paper
sandwiches.
Life Technologies makes blotting easier by providing a variety
of precut, preassembled membrane/filter-paper sandwiches
for mini and midi/E-PAGE™ gels (Figure 10). A protein’s
properties (i.e., charge, hydrophobicity, etc.) affect its ability to
bind to membrane surfaces. Finding the right membrane may
require experimenting with your specific protein on different
membranes.
For more information about Life Technologies western analysis
products, go to lifetechnologies.com/iblot
Figure 9. The iBlot® 7-Minute Blotting System.
1
2
3
A
4
1
2
3
4
B
Invitrolon™ PVDF
Other 0.45 µm PVDF
Figure 10. High signal achieved on Invitrolon™ PVDF. A 53 kDa
protein containing a c-myc epitope was transferred to (A) Invitrolon™
PVDF and (B) another manufacturer’s 0.45 µm PVDF membrane.
Both PVDF membranes were probed with a 1:500 dilution of mouse
anti-myc antibody, then developed with the WesternBreeze™
Chemiluminescent Anti-Mouse Kit. Blots shown here are 2 min
exposures on X-ray film. Lanes 1–4: 2 ng, 1 ng, 0.5 ng, and 0.2 ng of
protein.
15
Product
Quantity
Cat. No.
1 unit
IB1001
1 unit
IB1001EU
1 unit
IB1001UK
iBlot Transfer Stack, Regular (Nitrocellulose)
10 sets/box
IB3010-01
iBlot® Transfer Stack, Mini (Nitrocellulose)†
10 sets/box
IB3010-02
10 sets/box
IB4010-01
iBlot Transfer Stack PVDF, Mini
10 sets/box
IB4010-02
Blotting Roller, 8.6 cm wide
1 unit
LC2100
Incubation Tray, 10 x 14 x 3 cm (L x W x D)
8 trays and lids/pkg
LC2102
Semi-Dry Blotter
1 unit
SD1000
Blotting Filter Papers, 2.5 mm thickness, 8.6 x 13.5 cm
50 filter papers
LC2008
Blotting Filter Papers, 2.5 mm thickness, 7.5 x 8.4 cm
50 filter papers
LC2010
20 sets/pkg
LC2002
Invitrolon PVDF/Filter Paper Sandwich, 0.45 µm pore size, 8.5 x 13.5 cm
16 sets/pkg
LC2007
Nitrocellulose/Filter Paper Sandwich, 0.45 µm pore size, 8.5 x 13.5 cm
16 sets/pkg
LC2006
Nitrocellulose/Filter Paper Sandwich, 0.2 µm pore size, 8.5 x 13.5 cm
16 sets/pkg
LC2009
1 kit
BN3001
Western blotting
Dry electroblotting
iBlot® Gel Transfer Device
®
iBlot Gel Transfer Device (EU)
®
iBlot Gel Transfer Device (UK)
®
†
®
†
iBlot Transfer Stack PVDF, Regular
®
†
Efficient semi-dry blotting
™
Invitrolon PVDF/Filter Paper Sandwich, 0.45 µm pore size, 8.3 x 7.3 cm
™
™
®
NativePure Lentiviral Gateway Vector Kit
Precut blotting membranes
Product
Applications
Size
No. of membrane/filterpaper sandwiches
Cat. No.
Nitrocellulose,
0.2 µm pore size
Western transfers, solid-phase
assay systems, amino acid analysis
8.3 x 7.3 cm (mini)
20
LC2000
8.5 x 13.5 cm (midi)
16
LC2009
8.3 x 7.3 cm (mini)
20
LC2001
8.5 x 13.5 cm(midi)
16
LC2006
8.3 x 7.3 cm (mini)
20
LC2005
8.5 x 13.5 cm (midi)
16
LC2007
Nitrocellulose,
0.45 µm pore size
™
Invitrolon PVDF,
0.45 µm pore size
†
Western transfers, solid-phase
assay systems, amino acid analysis
Western transfers, solid-phase
assay systems, amino acid analysis, reprobing
PVDF,
0.2 µm pore size
Western transfers, solid-phase
assay systems, amino acid analysis, reprobing
8.3 x 7.3 cm (mini)
20
LC2002
Nylon,
0.45 µm pore size
Southern, northern, and western transfers,
solid-phase immobilization, dry chemistry test strips,
enzyme immobilization, gene probe assays
8.3 x 7.3 cm (mini)
20
LC2003
Each iBlot® bottom stack includes an integrated 0.2 μm nitrocellulose or polyvinylidene difluoride (PVDF) membrane for protein immobilization.
16
®
BenchPro 4100 Western Processing System
Just push “run” and walk away
ǩ Reduces tedious hands-on work and human errors
ǩ Minimizes cross-contamination, and no clean-up
ǩ Automate without protocol changes
ǩ Compatible with all chemiluminescent, chromogenic, and fluorescent immunodetection reagents and protocols
The BenchPro® 4100 Western Processing System (Figure 11) helps consistently and accurately deliver the correct volumes of
solutions to the membrane at precise times. This automation removes the tedious and repetitive liquid handling steps associated
with manual western blot processing. Western blots processed using the BenchPro® 4100 system minimize human processing
errors, so you get greater experimental reproducibility and data consistency.
The BenchPro® 4100 Western Processing System is designed to generate familiar results the first time and consistent results
on subsequent runs. It is compatible with all chemiluminescent, chromogenic, and fluorescent immunodetection reagents and
protocols (Figure 12). The BenchPro® 4100 system dramatically reduces the need for protocol optimization and allows you to
perform immunoblotting your way, right away.
For more information, go to lifetechnologies.com /benchpro4100
1
2
3
4
A
Figure 11. BenchPro® 4100 Western Processing System.
Product
Cat. No.
1 each
WP0001
BenchPro 4100 Western Card
10 cards
WP1001
BenchPro® 4100 Reagent Vials
50 vials
WP3001
®
2
3
4
Figure 12. Manual processing vs. BenchPro® 4100 processing of
western blots. (A) Manual processing; (B) BenchPro® 4100 system
processing. Lane 1: 8 µL of a 1:10 dilution of MagicMark™ XP Standard;
lane 2: 50 ng BSA; lane 3: 25 ng BSA; lane 4: 10 ng BSA. Proteins
were detected using rabbit anti-BSA antibody and WesternBreeze™
Chemiluminescent Kit—Anti-Rabbit. Detection substrate was added to
both membranes, and the blots were imaged at the same time.
Quantity
BenchPro® 4100 Card Processing Station
1
B
17
®
Novex western blot detection kits
Life Technologies offers a broad range of reagents and kits for western blot detection. Whether you are using chromogenic,
fluorescent, or chemiluminescence detection systems, we have the solutions you need.
iBlot® Western Detection Kits
ǩ Fast—complete western detection, typically in less than 25 minutes when used with the iBlot® 7-Minute Blotting System
ǩ Flexible—works with mini and midi gels
ǩ Easy optimization—allows the use of different conditions for different sections of the blot
The iBlot® Western Detection Kits (Figure 13) consist of iBlot® detection stacks and iBlot® detection reagents. The kits are
available with anti-mouse or anti-rabbit secondary antibodies and are compatible with chemiluminescent and chromogenic
detection. The iBlot® Western Detection Kits offer comparable or better sensitivity than the conventional protocols for the
majority of the antibody-antigen pairs, while offering significant time savings.
1
2
3
4
5
A
1
2
3
B
4
5
Figure 13. Comparison of an iBlot® Western Detection Kit to
a WesternBreeze™ kit. Proteins from an SW480 (human colon
adenocarcinoma cell line) lysate were transferred using the iBlot®
7-Minute Blotting System. Mouse anti-p53 antibody was used
as the primary antibody. Detection was performed with (A) the
WesternBreeze™ Chemiluminescent Kit—Anti-Mouse or (B) the iBlot®
Western Detection, Chemiluminescent Kit (Anti-Mouse). The iBlot®
western detection kit detected the proteins with sensitivity comparable
to the WesternBreeze™ kit.
Product
Quantity
Cat. No.
iBlot® Gel Transfer Device
1 device
IB1001
®
10 reactions
IB7110-01
®
iBlot Western Detection Chemiluminescent Kit (Anti-Mouse)—Mini, 10 Pak
10 reactions
IB7110-02
iBlot® Western Detection Chemiluminescent Kit (Anti-Rabbit)—Regular, 10 Pak
10 reactions
IB7210-01
iBlot® Western Detection Chemiluminescent Kit (Anti–Rabbit)—Mini, 10 Pak
iBlot Western Detection Chemiluminescent Kit (Anti-Mouse)—Regular, 10 Pak
10 reactions
IB7210-02
®
10 reactions
IB7310-01
®
10 reactions
IB7310-02
®
10 reactions
IB7410-01
®
10 reactions
IB7410-02
®
10 reactions
IB7010-01
®
10 reactions
IB6310-02
iBlot Western Detection Chromogenic Kit (Anti-Mouse)—Regular, 10 Pak
iBlot Western Detection Chromogenic Kit (Anti-Mouse)—Mini, 10 Pak
iBlot Western Detection Chromogenic Kit (Anti-Rabbit)—Regular, 10 Pak
iBlot Western Detection Chromogenic Kit (Anti–Rabbit)—Mini, 10 Pak
IBlot Western Detection Stacks (Regular), 10 Pak
iBlot Western Detection Stacks (Mini), 10 Pak
18
Additional Novex® western detection kits
WesternBreeze™
Chromogenic Kit
Product
Strong, long-lasting signal
Application
WesternBreeze™
Chemiluminescent Kit
ECL
Chemiluminescent Kit
Highly sensitive detection
®
WesternDot™ Blotting Kit
Using horseradish
Straightforward, high-
with ready-to-use BCIP/
with ready-to-use CDP-Star
peroxidase, low-picogram
sensitivity detection of
NBT substrate for alkaline
chemiluminescent substrate
levels of substrate can
proteins using fluorescent
phosphatase
for alkaline phosphatase
be detected with the ECL
Qdot® 625 nanocrystals
Chemiluminescent Kit
combined with high-affinity
streptavidin-biotin binding
reaction
Sensitivity
Low-picogram
High-femtogram
High-femtogram
Low-picogram
Emission duration
Months to years
Days
Hours
Days to months
Additional
equipment required
None Optional:
Autorad/x-ray film and
Autorad/X-ray film and
UV transilluminator or
white-light camera or
developer
developer
UV illuminator
scanner
Scanner
Scanner
Digital camera with orange/
Luminescent imager
Luminescent imager
red filter
Laser scanner
Product
Quantity
Cat. No.
Chromogenic and chemiluminescent immunodetection
WesternBreeze™ Chromogenic Kit—Anti-Mouse
1 kit
WB7103
™
1 kit
WB7105
™
WesternBreeze Chromogenic Kit—Anti-Goat
1 kit
WB7107
WesternBreeze™ Chemiluminescent Kit—Anti-Mouse
1 kit
WB7104
WesternBreeze™ Chemiluminescent Kit—Anti-Rabbit
WesternBreeze Chromogenic Kit—Anti-Rabbit
1 kit
WB7106
™
1 kit
WB7108
™
80 mL each
WB7050
™
2 x 100 mL
WB7003
250 µL
LC5602
1 kit
WP20005
1 kit
W10132
1 kit
W10142
WesternBreeze Chemiluminescent Kit—Anti-Goat
WesternBreeze Blocker/Diluent (part A and B)
WesternBreeze Wash Solution (16X)
Easy molecular weight estimation directly on western blots
MagicMark™ XP Western Protein Standard
®
Novex ECL Chemiluminescent Kits
™
WesternDot Blotting Kit
WesternDot™ 625 Goat Anti-Mouse Western Blot Kit
™
WesternDot 625 Goat Anti-Rabbit Western Blot Kit
19
To learn more about the complete protein separation, blotting, and detection solutions provided in Novex®
protein analysis systems, go to lifetechnologies.com/novex
For research use only. Not intended for any animal or human therapeutic or diagnostic use. ©2012 Life Technologies Corporation. All rights
reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. Coomassie is a
trademark of BASF Aktiengesellschaft. Criterion is a trademark of Bio-Rad Laboratories, Inc. CO01799 0312
lifetechnologies.com