Bolt gels let you see western blot bands more clearly than ever before
Invitrogen Bolt Bis-Tris Plus Gels are optimized gels for western blot analysis. They offer high sample resolution and better sample integrity with easier-to-load, larger capacity wells.
Bolt Bis-Tris Plus Gels are optimized gels for western blot analysis. Some features include:
- Superior band quality and band volume—Invitrogen Bolt Bis-Tris Plus chemistry is designed to deliver sharp, straight bands with higher band volume.
- Better protein resolution—more protein bands are detected due to 10% greater resolving distance and optimized gradient format.
- Preserved protein integrity—the legendary neutral-pH formulation minimizes protein modifications.
- Superior lot-to-lot consistency—our best-in-class manufacturing gives you highly reproducible gel performance, with a CV of only 2% for Rf resolution values.
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 the iBind processing protocols with 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 (Jackson ImmunoResearch). Detection was performed with Novex ECL HRP Substrate.
More accurate protein molecular weight assignments
Bolt gels are precast SDS-PAGE gels designed to give optimal resolution, band quality, and sensitivity for all your western blot analyses. Bolt gels are 10% longer than 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 Bio-Rad’s gels, which may present poorly-resolved, diffuse or streaky bands. Besides providing you with more accurate protein molecular weight assignments, Bolt gels offer 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 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. 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 Bio-Rad’s gels. The western workflow requires significant gel handling, so we developed Bolt gels to be stronger than other commercially available gels in order to minimize gel tearing.
More sample per well
Our unique, innovative wedge-shaped 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 a compromise in quality. With our 7-minute Invitrogen iBlot 2 western blot transfer process, now you can be western-ready in as little as 42 minutes without sacrificing quality.
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.
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 Bio-Rad’s tris-glycine gradient gels (B). Western detection was performed for (A) and (B) with Invitrogen 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 seconds. Chromogenic detection was performed for gels (C) and (D). Lane 1: blend of 6 purified proteins; lanes 2–4, 6–7: 250 ng of 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: Invitrogen MagicMark XP Western Protein Standard.
Bolt band integrity
Bolt gels preserve protein integrity with neutral-pH gel chemistry. This formulation minimizes protein modifications during electrophoresis.
Bolt 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 compared to other commercially available precast gels. 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. (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). The resolution of the two bands is superior on the Bolt gel.
The Bolt Bis-Tris Plus chemistry and optimized well dimensions are designed to 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 Bio-Rad’s gels. Western blots of dilution series of GST fusion proteins loaded on Bolt gels show even, stepwise gradations, whereas Bio-Rad’s gels show less even gradation of the same dilution series. In addition, the overall western signal detected on the blot of the Bolt gel, as defined by band volume, is higher than on the blot of Bio-Rad’s gel.
For Research Use Only. Not for use in diagnostic procedures.