First Isolation of Full-Length BRCA2 Protein Enabled by Bio-Rad Mini-PROTEAN TGX Precast Gels
Hercules, CA — Aug. 27, 2010 — Dr Ryan Jensen of the University of California, Davis and colleagues reported the first purification of full-length BRCA2 protein in a paper published online August 22 in the journal Nature. To follow the purification process and to analyze the protein's biochemistry, Jensen used Bio-Rad's Mini-PROTEAN TGX (Tris-Glycine eXtended) precast gels.
Proper study of the product of the BRCA2 gene, mutations of which lead to breast and ovarian cancers, has eluded research groups for years due to challenges preventing its purification. The protein is large (3,418 amino acids), has low expression levels, and a tendency to degrade.
Like many researchers, prior to using Mini-PROTEAN TGX gels Jensen used his own handcast polyacrylamide gels to trace the purification of BRCA2. During many of these attempts, the higher molecular weight proteins —including BRCA2 — disappeared.
Once Jensen began using the Mini-PROTEAN TGX gels, he consistently observed "tight, crisp" bands corresponding to BRCA2.
"When I hand poured my gels to analyze the purification of BRCA2, I wasn't always confident that it would show up," Jensen said. "With Bio-Rad's TGX gels, I could count on the fact that BRCA2 would show up every time."
The Mini-PROTEAN TGX gels are based on a modification of the Laemmli buffer system, which increases gel matrix stability and performance over time. This modification extends the gel shelf life to more than 12 months and offers superior resolution and reproducibility. Researchers can run gels to completion in as little as 12 minutes.
Jensen's absolute confidence in Bio-Rad's gels was especially important due to the small amount of protein at his disposal and the fact that other labs around the world were racing to purify the same protein and publish their results. Jensen appreciated the time savings and convenience of not having to pour gels, allowing him to focus on his research.
For Jensen, an additional advantage over other precast gels was that he could use the standard and inexpensive Tris/Glycine/SDS buffers for sample loading and running. Other precast gels require customized buffers.
"I think there's a pervasive feeling among researchers that their own hand-poured gels have better resolution, but I would argue the opposite," said Jensen. "The TGX gels provide a facile solution for producing publication quality figures."
Jensen was able to perform downstream techniques such as Western blotting with reproducible results. Jensen also remarked that using Bio-Rad's precast gels prevented user contamination that could potentially interfere with mass spectrometric analysis when using handcast gels.
For product details or to order Mini-PROTEAN TGX precast gels, visit http://www.miniprotean.com.
Posted: August 2010