Invitrogen precast Bis-Tris gels

NuPAGE Bis-Tris Gels and Bolt Bis-Tris Plus Gels are high-performance precast polyacrylamide gels developed to provide optimal separation of a wide range of protein sizes under denaturing conditions. Bis-Tris chemistry delivers consistent gel performance with a neutral pH environment that minimizes protein modifications. NuPAGE and Bolt gels are a good choice before downstream transfer and sensitive western blot analysis and any other technique where protein integrity is crucial. The unique WedgeWell design of the Bolt Bis-Tris Plus gel allows loading of up to 2X more sample volume than other precast gels. Bis-Tris chemistry in NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels also promotes stability of the polyacrylamide gel matrix, enabling room temperature storage, which helps save on the need for refrigerator space.

NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gel specifications

  NuPAGE Bis-Tris Bolt Bis-Tris Plus
Select when Need flexibility in well and gel format Working with dilute samples or need to load larger sample volumes, or when faster run times are needed
Available gel sizes Mini: 8 cm x 8 cm (1.0 or 1.5 mm thick)
Midi: 8 cm x 13 cm (1.0 mm thick)
Mini: 8 cm x 8 cm (1.0 mm thick)
Storage conditions 4–25°C
Shelf life 12 months 16 months
High lot-to-lot consistency Coefficient of variation (CV) of only 2% for Rf values (migration) Coefficient of variation (CV) of only 2% for Rf values (migration)
Recommended sample buffer NuPAGE LDS Sample Buffer Bolt LDS sample buffer
Recommended running buffers NuPAGE MES SDS or NuPAGE MOPS SDS Running Buffer Bolt MES SDS or Bolt MOPS SDS running buffer
Run time 35 min (MES buffer)
50 min (MOPS Buffer)
20 min (MES buffer)
35 min (MOPS buffer)
Recommended transfer buffer NuPAGE Transfer Buffer Bolt Transfer Buffer
Gel chemistry Bis-Tris
Available polyacrylamide concentrations 8%, 10%, 12%, 4–12%
Separation range 15 kDA to 260 kDa (MOPS buffer)
3.5 kDA to 160 kDa (MES buffer)
For use with (equipment) mini gels Mini Gel Tank or XCell SureLock Mini-Cell
For use with (equipment) midi gels SureLock Tandem Midi Gel Tank, Invitrogen XCell4 SureLock Midi-Cell or Bio-Rad Criterion (with adapters only) Not available in midi size
Mode of separation Molecular weight
Application SDS-PAGE
Available Wells configurations* Mini: 1, 9, 10, 12, 15, 17, 2D-well, IPG well
Midi: 12+2, 20, 26 wells
WedgeWell format: 10, 12, 15 17 well (load up to 2X sample volume per well)

*Not all percentages are available in every well type

Invitrogen Bis-Tris Gel Highlights

Sharp, straight bands

Neutral-pH buffers in NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels deliver sharp straight bands. During separation, the gels operate close to pH 7. In the Laemmli system (Tris-glycine), the gel is run at basic pH (pH ~9.5). At high pH the residual unpolymerized acrylamide can react with cysteine and lysine residues of the proteins being separated. This may affect downstream analysis such as western blot transfer. At the neutral pH of NuPAGE Bis-Tris and Bolt Bis-Tris Gels, these reactions take place several orders of magnitude more slowly than at the basic pH of Tris-glycine gels, resulting in sharper bands and better resolution.

NuPAGE Bis-Tris gels provide sharp straight bands compared to competitor tris glycine gels

Protein separation comparing NuPAGE Bis-Tris Gel and traditional tris-glycine gel. The samples listed below were run on (A) a NuPAGE 4–12% Bis-Tris Gel in Invitrogen NuPAGE MES SDS Running Buffer or (B) another manufacturer’s 4–20% tris-glycine gel. Note the high resolution of the sample bands in the NuPAGE protein gel. The poorly resolved “fuzzy” bands seen in the other manufacturer’s gel are a result of reoxidation of some disulfide bonds within the sample, leading to slight changes in migration rates. Lanes 1, 6, 7, 10: Invitrogen Mark12 Unstained Standard; lane 2: high protein load (4 proteins, 6 µg/protein); lane 3: Broad Range Standard; lanes 4, 5: E. coli extract; lane 8: E-PAGE SeeBlue Pre-stained Standard; lane 9: Invitrogen MultiMark Multi-Colored Protein Standard.

Better sample integrity

Sample preparation is a crucial step in successful protein analysis. The pH of the traditional Laemmli-style sample buffer changes from 6.8 to 5.2 when heated at 100°C. This lower pH is known to induce aspartyl-prolyl (Asp-Pro) peptide bond cleavage, which leads to protein degradation. By maintaining a >7.0 pH environment, the Invitrogen NuPAGE LDS Sample Buffer and Bolt LDS Sample Buffer preserve protein integrity by minimizing this Asp-Pro cleavage. As seen below, the sample integrity is maintained throughout electrophoresis with the Invitrogen NuPAGE system; protein degradation is seen in samples prepared with Laemmli (Tris-glycine) sample buffer. The Invitrogen NuPAGE Antioxidant running buffer additive greatly minimizes protein oxidation during electrophoresis and keeps reduced protein bands sharp and clear.

NuPAGE gel system provides better sample integrity than the traditional Laemmli (tris-glycine) gel system

High sample integrity. Integrity of samples is maintained throughout electrophoresis with the NuPAGE Bis-Tris Gel System (left), compared to samples prepared with Laemmli (tris-glycine) sample buffer (right).

Bolt Bis-Tris Plus Gel high capacity WedgeWell sample wells

The unique wedge-shaped well in every Bolt gel provides higher loading capacity, so you can load up to twice as much protein sample in every well.

Well size Recommended loading volume Maximum loading volume Maximum protein load
10-well 40 µL 60 µL 0.5 µg/band
12-well 30 µL 45 µL 0.4 µg/band
15-well 20 µL 35 µL 0.25 µg/band
17-well 15 µL 30 µL 0.2 µg/band

Detect proteins even in dilute samples or measure expression of low-abundance proteins. With more room to load your sample, you no longer have to worry about sample spillover and cross-well contamination on your western blots. Gel loading is also easier with no need to use special gel-loading pipette tips.

NuPAGE Bis-Tris Gels and Bolt Bis-Tris Plus Gels

NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels are precast polyacrylamide gels designed to give optimal separation for a wide range of molecular weight proteins during gel electrophoresis under denaturing conditions. The Bis-Tris gel chemistry provides a neutral-pH environment that minimizes protein modifications, resulting in sharper bands without typical gel "smiling". Additionally, preserving protein integrity becomes particularly important when separating low-abundance proteins.

NuPAGE Bis-Tris gels are offered as mini and midi gels, and Bolt Bis-Tris Plus gels are offered as mini gels. Both NuPAGE Bis-Tris gels and Bolt Bis-Tris Plus gels are ideal for western blot analysis and applications such as sequencing and mass spectrometry where protein integrity is crucial and higher sensitivity is required. The unique WedgeWell design of the Bolt Bis-Tris Plus gel sample well allows loading of up to 2x more sample volume than other precast gels.

Features of the NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels include:

  • Better protein integrity—optimized sample preparation process preserves your proteins
  • Wide ranges of molecular weight separation—selecting the right gel and running buffer helps to achieve the optimal separation of your proteins
  • Faster run times—separation of your proteins can be attained in 22-35 minutes
  • Longer shelf life—NuPAGE Bis-Tris and Bolt Bis-Tris Plus gels can be stored for at least 12 months and 16 months at room temperature, respectively
  • High capacity wells of Bolt Bis-Tris Plus Gels—wedge-shaped wells makes sample loading easier (no specialized pipette tips required) and allow loads of up to two times higher sample volume

NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels are formulated for denaturing gel electrophoresis applications. For optimal sample preparation, we recommend using buffers containing SDS. The gels can be run using MES SDS running buffer to better resolve smaller proteins (1–200 kDa) and MOPS SDS running buffer to resolve medium- to large-size proteins (14–260 kDa).

Bis-Tris Gels use a Bis-Tris discontinuous buffer system involving three ions:

  • Chloride (–) is from the gel buffer and serves as a leading ion due to its high affinity to the anode relative to other anions in the system. The gel buffer ions are bis-tris (+) and Cl (–) (pH 6.4).
  • MES or MOPS (–) serves as the trailing ion in the running buffer. The running buffer ions are tris (+), MOPS/MES (–) and dodecylsuflate (pH 7.3–7.7).

Bis-Tris (+) is the common ion present in the gel buffer and running buffer. The combination of lower pH gel buffer (pH 6.4) and the running buffer (pH 7.3–7.7) results in a significantly lower operating pH of 7 during electrophoresis.

Get started with Protein gel welcome packs

Protein gel welcome packs

Protein gel welcome packs are a cost-effective way to start using Invitrogen precast protein gels for the first time. Protein gel welcome packs are available for each of the protein gel chemistries and percentages and come with mini gels, buffers, ladders, and a Mini Gel Tank to provide you with great savings over the cost of the individual components.

Optimized performance of NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels

Unlike traditional Tris-glycine gels, NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels are Bis-Tris HCl buffered (pH 6.4) and have an operating pH of about 7.0. This neutral pH paired with a unique, gentle sample preparation protocol means your protein samples always reside in mild, nonacidic conditions, preserving the integrity of the proteins and minimizing protein modifications. This helps to ensure highly sensitive, accurate western blot results every time.

Western blot analysis using a Bolt Bis-Tris Plus gel compared to tris-glycine gels

Greater sensitivity with Bolt Bis-Tris Plus gels. Total cell extracts from A431 cells were transferred to NC and PVDF membranes from a 4–12% Bolt Bis-Tris Plus gel, and 4–20% Tris-glycine precast gel using the iBlot 2 Gel Transfer Device. The cells were treated with 100 ng/mL of human epidermal growth factor (hEGF) to up-regulate expression of the phospho-EGF receptor. The protein loads of the cell extracts ranged from 20 μg to 1.2 μg of extract. The blots were processed on the iBind Western System using a 1:200 dilution of Phospho-EGF Receptor (Tyr1068) (1H12) Mouse mAb (Cell Signaling Technology) and a 1:2,000 dilution of anti-mouse HRP secondary antibody.

Band integrity

NuPAGE and Bolt gels preserve protein integrity with neutral-pH gel chemistry. This formulation minimizes protein modifications during electrophoresis.

A western blot of a Bolt Bis-Tris Plus gel shows clean, sharp protein signals corresponding to only full-length proteins, whereas a western blot of a Bio-Rad TGX gel shows multiple low molecular weight degradation products. Protein kinases implicated in cancer (IKKϐ, EPHB3, HCK, MAPK14, FLT1 and DDR2) were analyzed on a Bolt Bis-Tris Plus Gel and a Bio-Rad TGX tris-glycine gel. The purified kinases (50 ng each), along with MagicMark protein standard and purified recombinant GST protein, were loaded on a 10-well, 4–12% Bolt gel and a 10-well, 4–20% Bio-Rad TGX gel. The samples were separated and transferred to 0.45-μm PVDF membranes using the respective manufacturers’ protocols. Immunodetection was performed using an anti-GST antibody and WesternBreeze chemiluminescence detection.

Band resolution

NuPAGE and Bolt Bis-Tris Plus Gels have better resolving power than Bio-Rad’s gels. With 10% greater resolving distance, optimized gel chemistry, and gradient format, you can see more protein bands resolved on a Bolt gel. In experimental analysis, you can see two separate bands for the insulin B chain and insulin A chain, which have similar molecular weights. The protein bands on the Bolt gel appear straight and the signal intensities are even across the entire bands, whereas the protein bands on Bio-Rad’s gel are uneven and of poor quality.

Bolt Bis-Tris Gel band resolution—comparison of Bio-Rad's stained protein gels

Bolt Bis-Tris Gel band resolution—comparison of Bio-Rad's stained protein gels. (A) Bolt Bis-Tris gradient gel stained with Invitrogen SimplyBlue SafeStain. (B) Bio-Rad's tris-glycine gradient gel stained with competitor’s stain. (C) Enlargement of a section of lane 5 from the two gels shows the insulin B chain (3.5 kDa) and insulin A chain (2.5 kDa).

Use the protein gel selection guide to find the best mini or midi gel for your protein electrophoresis experiment.

Find the best Invitrogen mini or midi gel to replace your current precast gel from another supplier using the protein gel conversion tool.

Currently using Tris-glycine or Tricine gels? Use the NuPAGE Gel conversion guide below to find the appropriate NuPAGE gel and corresponding optimized buffer.

NuPAGE Gel conversion guide

If you are using: Then we recommend NuPAGE gel: Buffer system
Tris-Glycine gel
4% NuPAGE 3-8% Tris-Acetate Tris-Acetate
6% NuPAGE 3-8% Tris-Acetate Tris-Acetate
10% NuPAGE 8% Bis-Tris MOPS
NuPAGE 10% Bis-Tris MOPS
12% NuPAGE 10% Bis-Tris MOPS
14% NuPAGE 12% Bis-Tris MOPS
16% NuPAGE 12% Bis-Tris MES
18% NuPAGE 12% Bis-Tris MES
4-12% NuPAGE 3-8% Tris-Acetate Tris-Acetate
NuPAGE 4-12% Bis-Tris MOPS
4-20% NuPAGE 4-12% Bis-Tris MES
8-16% NuPAGE 4-12% Bis-Tris MOPS
10-20% NuPAGE 12% Bis-Tris MOPS
Tricine gel
10% NuPAGE 8% Bis-Tris MES
NuPAGE 10% Bis-Tris MES
16% NuPAGE 4-12% Bis-Tris MES
10-20% NuPAGE 4-12% Bis-Tris MES

Download the protein gel guide for a printable gel migration chart.

Migration chart for NuPAGE Bis-Tris Gels
Chart showing the migration pattern of a collection of standard proteins when separated on different Bolt Bis-Tris Plus Gels

NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gel specifications

  NuPAGE Bis-Tris Bolt Bis-Tris Plus
Select when Need flexibility in well and gel format Working with dilute samples or need to load larger sample volumes, or when faster run times are needed
Available gel sizes Mini: 8 cm x 8 cm (1.0 or 1.5 mm thick)
Midi: 8 cm x 13 cm (1.0 mm thick)
Mini: 8 cm x 8 cm (1.0 mm thick)
Storage conditions 4–25°C
Shelf life 12 months 16 months
High lot-to-lot consistency Coefficient of variation (CV) of only 2% for Rf values (migration) Coefficient of variation (CV) of only 2% for Rf values (migration)
Recommended sample buffer NuPAGE LDS Sample Buffer Bolt LDS sample buffer
Recommended running buffers NuPAGE MES SDS or NuPAGE MOPS SDS Running Buffer Bolt MES SDS or Bolt MOPS SDS running buffer
Run time 35 min (MES buffer)
50 min (MOPS Buffer)
20 min (MES buffer)
35 min (MOPS buffer)
Recommended transfer buffer NuPAGE Transfer Buffer Bolt Transfer Buffer
Gel chemistry Bis-Tris
Available polyacrylamide concentrations 8%, 10%, 12%, 4–12%
Separation range 15 kDA to 260 kDa (MOPS buffer)
3.5 kDA to 160 kDa (MES buffer)
For use with (equipment) mini gels Mini Gel Tank or XCell SureLock Mini-Cell
For use with (equipment) midi gels SureLock Tandem Midi Gel Tank, Invitrogen XCell4 SureLock Midi-Cell or Bio-Rad Criterion (with adapters only) Not available in midi size
Mode of separation Molecular weight
Application SDS-PAGE
Available Wells configurations* Mini: 1, 9, 10, 12, 15, 17, 2D-well, IPG well
Midi: 12+2, 20, 26 wells
WedgeWell format: 10, 12, 15 17 well (load up to 2X sample volume per well)

*Not all percentages are available in every well type

Invitrogen Bis-Tris Gel Highlights

Sharp, straight bands

Neutral-pH buffers in NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels deliver sharp straight bands. During separation, the gels operate close to pH 7. In the Laemmli system (Tris-glycine), the gel is run at basic pH (pH ~9.5). At high pH the residual unpolymerized acrylamide can react with cysteine and lysine residues of the proteins being separated. This may affect downstream analysis such as western blot transfer. At the neutral pH of NuPAGE Bis-Tris and Bolt Bis-Tris Gels, these reactions take place several orders of magnitude more slowly than at the basic pH of Tris-glycine gels, resulting in sharper bands and better resolution.

NuPAGE Bis-Tris gels provide sharp straight bands compared to competitor tris glycine gels

Protein separation comparing NuPAGE Bis-Tris Gel and traditional tris-glycine gel. The samples listed below were run on (A) a NuPAGE 4–12% Bis-Tris Gel in Invitrogen NuPAGE MES SDS Running Buffer or (B) another manufacturer’s 4–20% tris-glycine gel. Note the high resolution of the sample bands in the NuPAGE protein gel. The poorly resolved “fuzzy” bands seen in the other manufacturer’s gel are a result of reoxidation of some disulfide bonds within the sample, leading to slight changes in migration rates. Lanes 1, 6, 7, 10: Invitrogen Mark12 Unstained Standard; lane 2: high protein load (4 proteins, 6 µg/protein); lane 3: Broad Range Standard; lanes 4, 5: E. coli extract; lane 8: E-PAGE SeeBlue Pre-stained Standard; lane 9: Invitrogen MultiMark Multi-Colored Protein Standard.

Better sample integrity

Sample preparation is a crucial step in successful protein analysis. The pH of the traditional Laemmli-style sample buffer changes from 6.8 to 5.2 when heated at 100°C. This lower pH is known to induce aspartyl-prolyl (Asp-Pro) peptide bond cleavage, which leads to protein degradation. By maintaining a >7.0 pH environment, the Invitrogen NuPAGE LDS Sample Buffer and Bolt LDS Sample Buffer preserve protein integrity by minimizing this Asp-Pro cleavage. As seen below, the sample integrity is maintained throughout electrophoresis with the Invitrogen NuPAGE system; protein degradation is seen in samples prepared with Laemmli (Tris-glycine) sample buffer. The Invitrogen NuPAGE Antioxidant running buffer additive greatly minimizes protein oxidation during electrophoresis and keeps reduced protein bands sharp and clear.

NuPAGE gel system provides better sample integrity than the traditional Laemmli (tris-glycine) gel system

High sample integrity. Integrity of samples is maintained throughout electrophoresis with the NuPAGE Bis-Tris Gel System (left), compared to samples prepared with Laemmli (tris-glycine) sample buffer (right).

Bolt Bis-Tris Plus Gel high capacity WedgeWell sample wells

The unique wedge-shaped well in every Bolt gel provides higher loading capacity, so you can load up to twice as much protein sample in every well.

Well size Recommended loading volume Maximum loading volume Maximum protein load
10-well 40 µL 60 µL 0.5 µg/band
12-well 30 µL 45 µL 0.4 µg/band
15-well 20 µL 35 µL 0.25 µg/band
17-well 15 µL 30 µL 0.2 µg/band

Detect proteins even in dilute samples or measure expression of low-abundance proteins. With more room to load your sample, you no longer have to worry about sample spillover and cross-well contamination on your western blots. Gel loading is also easier with no need to use special gel-loading pipette tips.

NuPAGE Bis-Tris Gels and Bolt Bis-Tris Plus Gels

NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels are precast polyacrylamide gels designed to give optimal separation for a wide range of molecular weight proteins during gel electrophoresis under denaturing conditions. The Bis-Tris gel chemistry provides a neutral-pH environment that minimizes protein modifications, resulting in sharper bands without typical gel "smiling". Additionally, preserving protein integrity becomes particularly important when separating low-abundance proteins.

NuPAGE Bis-Tris gels are offered as mini and midi gels, and Bolt Bis-Tris Plus gels are offered as mini gels. Both NuPAGE Bis-Tris gels and Bolt Bis-Tris Plus gels are ideal for western blot analysis and applications such as sequencing and mass spectrometry where protein integrity is crucial and higher sensitivity is required. The unique WedgeWell design of the Bolt Bis-Tris Plus gel sample well allows loading of up to 2x more sample volume than other precast gels.

Features of the NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels include:

  • Better protein integrity—optimized sample preparation process preserves your proteins
  • Wide ranges of molecular weight separation—selecting the right gel and running buffer helps to achieve the optimal separation of your proteins
  • Faster run times—separation of your proteins can be attained in 22-35 minutes
  • Longer shelf life—NuPAGE Bis-Tris and Bolt Bis-Tris Plus gels can be stored for at least 12 months and 16 months at room temperature, respectively
  • High capacity wells of Bolt Bis-Tris Plus Gels—wedge-shaped wells makes sample loading easier (no specialized pipette tips required) and allow loads of up to two times higher sample volume

NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels are formulated for denaturing gel electrophoresis applications. For optimal sample preparation, we recommend using buffers containing SDS. The gels can be run using MES SDS running buffer to better resolve smaller proteins (1–200 kDa) and MOPS SDS running buffer to resolve medium- to large-size proteins (14–260 kDa).

Bis-Tris Gels use a Bis-Tris discontinuous buffer system involving three ions:

  • Chloride (–) is from the gel buffer and serves as a leading ion due to its high affinity to the anode relative to other anions in the system. The gel buffer ions are bis-tris (+) and Cl (–) (pH 6.4).
  • MES or MOPS (–) serves as the trailing ion in the running buffer. The running buffer ions are tris (+), MOPS/MES (–) and dodecylsuflate (pH 7.3–7.7).

Bis-Tris (+) is the common ion present in the gel buffer and running buffer. The combination of lower pH gel buffer (pH 6.4) and the running buffer (pH 7.3–7.7) results in a significantly lower operating pH of 7 during electrophoresis.

Get started with Protein gel welcome packs

Protein gel welcome packs

Protein gel welcome packs are a cost-effective way to start using Invitrogen precast protein gels for the first time. Protein gel welcome packs are available for each of the protein gel chemistries and percentages and come with mini gels, buffers, ladders, and a Mini Gel Tank to provide you with great savings over the cost of the individual components.

Optimized performance of NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels

Unlike traditional Tris-glycine gels, NuPAGE Bis-Tris and Bolt Bis-Tris Plus Gels are Bis-Tris HCl buffered (pH 6.4) and have an operating pH of about 7.0. This neutral pH paired with a unique, gentle sample preparation protocol means your protein samples always reside in mild, nonacidic conditions, preserving the integrity of the proteins and minimizing protein modifications. This helps to ensure highly sensitive, accurate western blot results every time.

Western blot analysis using a Bolt Bis-Tris Plus gel compared to tris-glycine gels

Greater sensitivity with Bolt Bis-Tris Plus gels. Total cell extracts from A431 cells were transferred to NC and PVDF membranes from a 4–12% Bolt Bis-Tris Plus gel, and 4–20% Tris-glycine precast gel using the iBlot 2 Gel Transfer Device. The cells were treated with 100 ng/mL of human epidermal growth factor (hEGF) to up-regulate expression of the phospho-EGF receptor. The protein loads of the cell extracts ranged from 20 μg to 1.2 μg of extract. The blots were processed on the iBind Western System using a 1:200 dilution of Phospho-EGF Receptor (Tyr1068) (1H12) Mouse mAb (Cell Signaling Technology) and a 1:2,000 dilution of anti-mouse HRP secondary antibody.

Band integrity

NuPAGE and Bolt gels preserve protein integrity with neutral-pH gel chemistry. This formulation minimizes protein modifications during electrophoresis.

A western blot of a Bolt Bis-Tris Plus gel shows clean, sharp protein signals corresponding to only full-length proteins, whereas a western blot of a Bio-Rad TGX gel shows multiple low molecular weight degradation products. Protein kinases implicated in cancer (IKKϐ, EPHB3, HCK, MAPK14, FLT1 and DDR2) were analyzed on a Bolt Bis-Tris Plus Gel and a Bio-Rad TGX tris-glycine gel. The purified kinases (50 ng each), along with MagicMark protein standard and purified recombinant GST protein, were loaded on a 10-well, 4–12% Bolt gel and a 10-well, 4–20% Bio-Rad TGX gel. The samples were separated and transferred to 0.45-μm PVDF membranes using the respective manufacturers’ protocols. Immunodetection was performed using an anti-GST antibody and WesternBreeze chemiluminescence detection.

Band resolution

NuPAGE and Bolt Bis-Tris Plus Gels have better resolving power than Bio-Rad’s gels. With 10% greater resolving distance, optimized gel chemistry, and gradient format, you can see more protein bands resolved on a Bolt gel. In experimental analysis, you can see two separate bands for the insulin B chain and insulin A chain, which have similar molecular weights. The protein bands on the Bolt gel appear straight and the signal intensities are even across the entire bands, whereas the protein bands on Bio-Rad’s gel are uneven and of poor quality.

Bolt Bis-Tris Gel band resolution—comparison of Bio-Rad's stained protein gels

Bolt Bis-Tris Gel band resolution—comparison of Bio-Rad's stained protein gels. (A) Bolt Bis-Tris gradient gel stained with Invitrogen SimplyBlue SafeStain. (B) Bio-Rad's tris-glycine gradient gel stained with competitor’s stain. (C) Enlargement of a section of lane 5 from the two gels shows the insulin B chain (3.5 kDa) and insulin A chain (2.5 kDa).

Use the protein gel selection guide to find the best mini or midi gel for your protein electrophoresis experiment.

Find the best Invitrogen mini or midi gel to replace your current precast gel from another supplier using the protein gel conversion tool.

Currently using Tris-glycine or Tricine gels? Use the NuPAGE Gel conversion guide below to find the appropriate NuPAGE gel and corresponding optimized buffer.

NuPAGE Gel conversion guide

If you are using: Then we recommend NuPAGE gel: Buffer system
Tris-Glycine gel
4% NuPAGE 3-8% Tris-Acetate Tris-Acetate
6% NuPAGE 3-8% Tris-Acetate Tris-Acetate
10% NuPAGE 8% Bis-Tris MOPS
NuPAGE 10% Bis-Tris MOPS
12% NuPAGE 10% Bis-Tris MOPS
14% NuPAGE 12% Bis-Tris MOPS
16% NuPAGE 12% Bis-Tris MES
18% NuPAGE 12% Bis-Tris MES
4-12% NuPAGE 3-8% Tris-Acetate Tris-Acetate
NuPAGE 4-12% Bis-Tris MOPS
4-20% NuPAGE 4-12% Bis-Tris MES
8-16% NuPAGE 4-12% Bis-Tris MOPS
10-20% NuPAGE 12% Bis-Tris MOPS
Tricine gel
10% NuPAGE 8% Bis-Tris MES
NuPAGE 10% Bis-Tris MES
16% NuPAGE 4-12% Bis-Tris MES
10-20% NuPAGE 4-12% Bis-Tris MES

Download the protein gel guide for a printable gel migration chart.

Migration chart for NuPAGE Bis-Tris Gels
Chart showing the migration pattern of a collection of standard proteins when separated on different Bolt Bis-Tris Plus Gels
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