Bolt® Bis-Tris Plus Gels
Bolt® gels let you see western blot bands more clearly than ever before
Bolt® Bis-Tris Plus gels are precast polyacrylamide gels designed for optimal separation of a broad molecular weight range of proteins under denaturing conditions during gel electrophoresis. Bolt® Bis-Tris Plus gels are designed to deliver consistent gel performance and provide a neutral pH environment that minimizes protein modifications. Bolt® gels are ideal for Western blot transfer and analysis, and any other techniques where protein integrity is crucial.
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Choose the right Bolt® Bis-Tris Plus gel
|Number of wells||Gel percentage|
Checkout our newly optimized protocol: Bolt® Bis-Tris Gel Protocol
Welcome Pack: Bolt® Gels and iBlot® 2 Device for $2,395
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The Bolt® Welcome Packs provide all of the necessary Bolt® gels and buffers you will need to begin using the Bolt® Mini Gel Tank.
Bolt® gel migration chart
For optimal and total separation ranges for each specific gel percentage, view the gel migration chart.
Complete your Bolt® system with the innovative Bolt® Mini Blot Module that conveniently fits into the chambers of the Bolt® Mini Gel Tank.
Optimized western blot performance
More accurate protein molecular weight assignments
Bolt® gels are precast SDS-PAGE gels designed to give optimal resolution, band quality, and sensitivity for all of your western blot analyses. Bolt® gels are 10% longer than other competitors available gels, which results in higher resolving distance and therefore better band resolution. The pore size of the Bis-Tris Plus chemistry and the well-to-well spacing on Bolt® gels offers well-resolved, straight western blot bands with optimal band quality as compared to Competitor B’s gels that may present poorly resolved, diffuse, or streaky bands. Besides providing you with more accurate protein molecular weight assignments, Bolt® gels provide, overall, better-looking, publication-quality western blots.
Preserving the integrity of your proteins
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 are always in mild, nonacidic conditions, preserving the integrity of your proteins and minimizing protein modifications to help ensure highly sensitive, accurate western blots every time. Bolt® gels deliver best-in-class inter-lot and intra-lot reproducibility, with resolution (Rf) CV of only 2%, so you can have increased confidence in your gel performance every day.
Bolt® gels have been shown to be less fragile than Competitor B’s gels. The western workflow requires significant gel handling, so we developed Bolt® gels to be stronger than other commercially available gels so that gel tearing is minimized.
More sample per well
Our unique, innovative wedge wells enable up to two times the protein sample loads, so you get easy sample loading and no more spillover.
Bolt® gels are designed for speed without compromise in quality. With our 7-minute iBlot® western blot transfer, now you can be western-ready in as little as 42 minutes without sacrificing quality.
The Bolt® wedge well
Bolt® gels have a new unique wedge-shaped well, so you can load up to twice as much protein sample in every well. Now you can detect your 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 much easier and you no longer need to use special gel-loading pipet tips.
The wedge well is designed to provide optimal protein stacking at the start of the gel run, resulting in improved protein resolution, especially in the high molecular weight range. With higher sample volumes, the wedge well ultimately delivers better western sensitivity and allows for greater success with low-affinity western antibodies.
High sample volume capacity of Bolt® wedge wells. (Left) Increasing volumes (40–70 μL) of a fluorescent protein standard were loaded in every other lane of a Bolt® 4–12% Bis-Tris Plus 10-well gel. (Right) Increasing volumes (10–40 μL) of the same protein standard were loaded in every other lane of Competitor B'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® band quality
Bolt® Bis-Tris Plus gels are designed to deliver well-resolved, straight bands with optimal band quality as compared to other commercially available precast gels.
Western blot band quality with Bolt® precast gels. GST fusion proteins were loaded at 20 µL sample volume. Sharp, straight western bands are observed with Bolt® Bis-Tris gradient gels (A) as compared to smeared, poorly resolved, and uneven western signals on Competitor B’s Tris-glycine gradient gels (B). Western detection was performed for A and B with WesternBreeze™ chemiluminescent detection using a mouse anti-GST monoclonal antibody at 1:1,000 dilution; an LAS-1000 (FujiFilm) imager was used with an exposure time of 40 sec. Chromogenic detection was performed for gels (C) and (D). Lane 1: blend of 6 purified proteins; lanes 2–4, 6–7: 250 ng various GST fusion proteins; lane 5: mix of GST fusion proteins from lanes 4 and 6; lane 8: p53-GST; lane 9: GST; lane 10: MagicMark™ XP Western Protein Standard. Internal research data, May 2012.
Bolt® band resolution
Bolt® Bis-Tris Plus gels have better resolving power. With 10% greater resolving distance, optimized gel chemistry, and gradient format, you can see more protein bands resolved on a Bolt® gel compared to other commercially available precast gels. You can see two separate bands for the insulin B chain and 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 Competitor B’s gel are uneven and of poor quality.
Bolt® Bis-Tris gel band resolution. Comparison of Competitor B's stained protein gels. (A) Bolt® Bis-Tris gradient gel stained with SimplyBlue™ SafeStain. (B) Competitor B's Tris-glycine gradient gel stained with competitor 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). The resolution of the two bands is superior on the Bolt® gel.
The Bolt™ Bis-Tris Plus chemistry and optimized well dimensions deliver well-to-well reproducibility and band volume correspondence to sample load, therefore providing better quantitation capabilities on Western blots as compared to westerns of Competitor B’s gels. Western blots of dilution series of GST fusion proteins loaded on Bolt™ gels show even, stepwise gradations whereas Competitor B’s gels show less even gradation of the same dilution series. In addition, the overall western signal that is detected on the blot of the Bolt™ gel, as defined by band volume, is higher than on the blot of Competitor B’s gel
Western blot band volumes on Bolt™ gels vs. Bio-Rad® TGX™ gels. (A) Blot of a Bolt™ 4–12% gel. (B) Blot of a Bio-Rad® TGX™ 4–20% gel. Two GST fusion proteins as well as GST were spiked into E. coli lysate (0.6 mg per gel lane) to create series of decreasing, increasing, or constant amounts of these proteins across the gel. The spiked amounts of the two fusion proteins ranged from 40 ng to 0.2 ng per gel lane. Electrophoresis and wet transfers to PVDF membranes were performed using the materials, methods, equipment, and instructions of each manufacturer. WesternBreeze™ Chemiluminescence Detection Kits were used with an anti-GST antibody; images were acquired with an LAS-1000 (FujiFilm) system. (C) Band volumes measured on western blots of Bolt™ and Bio-Rad® TGX™ gels. Eight wells on each gel were loaded with 2 ng of GST. Band volumes were calculated based on the width, length, and intensity of the detected band. Internal research data, May 2012.
Bolt™ Bis-Tris Plus gels are available for resolving proteins in the range of 0.3–260 kDa, depending upon the acrylamide percentage of the gel and buffer system being used. Under denaturing conditions, use Bolt™ Bis-Tris Plus gels with MOPS SDS Running Buffer to resolve 14–260 kDa proteins, and Bolt™ Bis-Tris Plus gels with MES SDS Running Buffer to resolve 0.3–260 kDa proteins.
Resolve large molecules with low-percentage gels, and small molecules with high-percentage gels. If the molecular weight of the molecule is unknown, or the sample contains a wide range of molecules, use a gradient gel. Bolt™ Bis-Tris Plus gels do not contain SDS; however, they are designed for performing denaturing gel electrophoresis.
NOTE: Do not use Bolt™ Bis-Tris Plus gels with other gel electrophoresis systems. The gel cassettes with the new wedge-shaped wells are designed specifically for the Bolt™ Digital Electrophoresis System and are not compatible with other equipment.
Bolt® protein integrity
A western blot of a Bolt® 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. The blots were imaged using an LAS-1000 system (FujiFilm).
Recommended loading volumes
The recommended loading volumes and maximum protein load per band are provided in the following table. Note: to ensure the highest band resolution, do not exceed the recommended loading volume.
|Well type||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|
Bolt® Bis-Tris Plus gels are available in mini gel format with dimensions of 10 cm x 10.5 cm. The gels are 1.0 mm thick. Due to the wedge-well format that allows much higher sample loads, the gels do not require additional thickness. Bolt® Bis-Tris Plus gels come in multiple well formats.
NuPAGE® Bis-Tris gels are Bis-Tris HCl buffered (pH 6.4) and have an operating pH of about 7.0. All of the gels can be run using either Bolt® MES SDS or Bolt® MOPS SDS running buffer to obtain different separation ranges.
The Bolt® Bis-Tris Plus discontinuous buffer system involves three ions:
- Chloride (–) from the gel buffer serves as a leading ion due to its high affinity for the anode relative to other anions in the system. The gel buffer ions are Bis-Tris(+) and Cl– (pH 6.4).
- MES or MOPS (–) serve as the trailing ion in the running buffer. The running buffer ions are Tris+, MOPS–/MES–, and dodecyl sulfate– (pH 7.3–7.7).
- Bis-Tris (+) is the common ion present in the gel buffer and running buffer. The combination of the 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.