26-Well Protein Gel for High-Throughput Sample Analysis

The Scripps Research Institute
Immunology Microbiology
Postdoctoral Researcher

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Company:

Bio-Rad

Product Name:

Criterion TGX Stain-Free™

Catalog Number:

5678095

My research focuses on bacterial cell wall hydrolases, where I generate gene deletion mutants to understand their roles in cell growth and cell envelope biology. I also construct complementation plasmids expressing wild-type hydrolases and mutant derivatives to restore mutant phenotypes. I use the Criterion TGX Stain-Free™ 26-Well Protein Gels for high-throughput western blot analysis to compare protein expression across wild-type strains, hydrolase mutants, and complemented strains. The large number of wells makes these gels especially useful for analyzing multiple samples and biological replicates in a single experiment.

Experimental Design and Results Summary

Application

SDS-PAGE protein separation followed by Western blot

Starting Material

Bacterial cell lysates

Tips

The sample loading capacity is relatively low, so make sure your protein samples are sufficiently concentrated before loading the gel.

Results Summary

Using the Criterion TGX Stain-Free™ 26-Well Protein Gel, I was able to screen a large number of Enterococcus strains and complementation constructs in parallel by western blot. This high-throughput workflow revealed that some of my cell wall hydrolase mutant proteins were not expressed, whereas the wild-type protein and several mutant derivatives showed robust expression in Enterococcus. The ability to analyze many samples simultaneously made it much easier to identify functional constructs and compare protein expression across strains.

DOI or PMID #

N/A

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Summary

The Good

Careful loading is needed due to small wells.

The Bad

none

The Bottom Line

An excellent choice when many samples need to be analyzed in one experiment

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