Best Universal Blocking Buffer for Western Blot

January 27, 2023

BlueskyReddit
University of Texas Medical Branch Galveston

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioRad

Product Name:

EveryBlot Blocking Buffer

Catalog Number:

12010020

This is the best blocking buffer that I have had the pleasure of using for my western blots. I first started with 5% milk in TBST, but quickly realized that was not the best method to detect phosphorylated protein. I switched to 3% BSA in TBST which worked a lot better than milk, but still could use a bit of polishing. This is where my search ends, this blocking buffer is not only the cleanest blot I have ever gotten but I can produce results in half the time! This reagent is a winner.

Experimental Design and Results Summary

Application

Western Blot

Starting Material

Protein lysates

Tips

Noticed a bold, non-specific band at ~17kDA.

Results Summary

I am looking to detect the upregulation of a specific phosphorylated protein in the context of infectious disease. Using this blocking buffer, I was able to produce publication-quality blots that provide more insight into an ongoing project in my laboratory.

DOI or PMID #

N/A

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Summary

The Good

Saves a lot of time.

The Bad

Can only be purchased in 500ml portions which can get used up quickly.

The Bottom Line

I highly recommend this blocking buffer reagent, it is suitable for all types of proteins of interest and cuts down a 2-3 day western blot experiment to 1 day!

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