Bolt-Bis-Tris Plus gels

Bolt Bis-Tris Plus Gels are precast polyacrylamide gels developed to provide optimal separation of a wide range of protein sizes under denaturing conditions. Bolt Bis-Tris Plus Gels are designed to deliver consistent gel performance with a neutral pH environment that minimizes protein modifications. As such, Bolt gels are a good choice before downstream transfer and western blot analysis and any other technique where protein integrity is crucial.

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Bolt Bis-Tris Gel specifications

Available gel sizes Mini: 8 cm x 8 cm (1.0 mm thick)
Storage conditions Room temperature
Shelf life 16 months
Recommended sample buffer Bolt LDS sample buffer
Recommended running buffers Bolt MES SDS or Bolt MOPS SDS running buffer
Recommended transfer buffer Bolt transfer buffer
Gel chemistry Bis-Tris
Available polyacrylamide concentrations 8%, 10%, 12% and 4–12%
Separation range (denaturing) 14 kDA to 260 kDa (MOPS buffer)
2 kDa to 200 kDa (MES buffer)
For use with (equipment) Mini Gels Mini Gel Tank or XCell SureLock Mini-Cell
Mode of separation Molecular weight
Application SDS-PAGE
Well type 1D, Invitrogen WedgeWell format (load up to 60 µL per well)

High capacity wells- WedgeWell

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.

High sample volume capacity of Bolt Bis-Tris Plus gels. (A) Increasing volumes (40–70μL) of a fluorescent protein standard were loaded in every other lane of an Invitrogen Bolt 4–12% Bis-Tris Plus 10-well Gel. (B) Increasing volumes (10–40 μL) of the same protein standard were loaded in every other lane of Bio-Rad's 4–20% 10-well gel. Sample spillover and cross-well contamination is observed as signals from the sample loaded in the adjacent wells.

Bolt Bis-Tris Plus Gels are precast polyacrylamide gels that use a bis-tris discontinuous buffer system. Bis-Tris gel buffer system maintains proteins in a neutral-pH environment which preserves protein integrity by minimizing protein modifications. As such, Bolt gels are a good choice before downstream transfer and western blot analysis and any other technique where protein integrity is crucial.

Features and benefits of Bis-Tris Plus Gels:

  • Neutral pH environment– helps prevent protein modifications
  • High capacity wells– wedge-shaped wells makes sample loading easier (no specialized pipette tips required) and allow loads of up to two times higher sample volume.
  • Long shelf life– gels can be stored for 16 months at 4-25 °C
  • Fast run times– separate proteins in approximately 22-35 minutes

Bolt Bis-Tris Plus Gels are formulated for denaturing gel electrophoresis applications. For optimal sample preparation, use 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 discontinuous buffer system:

  • 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.

Optimized performance of Bolt Bis-Tris Plus Gels

Unlike traditional tris-glycine gels, 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

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

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).

Bolt Bis-Tris Plus migration chart

Bolt Bis-Tris Gel specifications

Available gel sizes Mini: 8 cm x 8 cm (1.0 mm thick)
Storage conditions Room temperature
Shelf life 16 months
Recommended sample buffer Bolt LDS sample buffer
Recommended running buffers Bolt MES SDS or Bolt MOPS SDS running buffer
Recommended transfer buffer Bolt transfer buffer
Gel chemistry Bis-Tris
Available polyacrylamide concentrations 8%, 10%, 12% and 4–12%
Separation range (denaturing) 14 kDA to 260 kDa (MOPS buffer)
2 kDa to 200 kDa (MES buffer)
For use with (equipment) Mini Gels Mini Gel Tank or XCell SureLock Mini-Cell
Mode of separation Molecular weight
Application SDS-PAGE
Well type 1D, Invitrogen WedgeWell format (load up to 60 µL per well)

High capacity wells- WedgeWell

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.

High sample volume capacity of Bolt Bis-Tris Plus gels. (A) Increasing volumes (40–70μL) of a fluorescent protein standard were loaded in every other lane of an Invitrogen Bolt 4–12% Bis-Tris Plus 10-well Gel. (B) Increasing volumes (10–40 μL) of the same protein standard were loaded in every other lane of Bio-Rad's 4–20% 10-well gel. Sample spillover and cross-well contamination is observed as signals from the sample loaded in the adjacent wells.

Bolt Bis-Tris Plus Gels are precast polyacrylamide gels that use a bis-tris discontinuous buffer system. Bis-Tris gel buffer system maintains proteins in a neutral-pH environment which preserves protein integrity by minimizing protein modifications. As such, Bolt gels are a good choice before downstream transfer and western blot analysis and any other technique where protein integrity is crucial.

Features and benefits of Bis-Tris Plus Gels:

  • Neutral pH environment– helps prevent protein modifications
  • High capacity wells– wedge-shaped wells makes sample loading easier (no specialized pipette tips required) and allow loads of up to two times higher sample volume.
  • Long shelf life– gels can be stored for 16 months at 4-25 °C
  • Fast run times– separate proteins in approximately 22-35 minutes

Bolt Bis-Tris Plus Gels are formulated for denaturing gel electrophoresis applications. For optimal sample preparation, use 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 discontinuous buffer system:

  • 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.

Optimized performance of Bolt Bis-Tris Plus Gels

Unlike traditional tris-glycine gels, 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

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

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).

Bolt Bis-Tris Plus migration chart
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