Simple and Cheap Beads for Immunoprecipitation From Santa Cruz

September 01, 2023

BlueskyReddit
Stanford University
Department of Pathology
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Santa Cruz

Product Name:

Protein A/G PLUS-Agarose

Catalog Number:

sc-2003

I regularly use Protein A/G agarose beads to perform immunoprecipitation experiments followed by Western blotting to detect my protein of interest.

Experimental Design and Results Summary

Application

Immunoprecipitation

Starting Material

Protein lysate from cells expressing Flag-tagged protein

Tips

Gently resuspend the beads and make sure equal volume of beads is added to the samples. The incubation times and the washings can be increased for optimised results.

Results Summary

The IP was performed according to the manufacturer's protocol. I used 1ug of Flag antibody for my protein of interest and corresponding IgG as control. To confirm pulldown of my protein, I performed western blot by boiling beads in SDS loading buffer.

DOI or PMID #

N/A

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Summary

The Good

Ease of use, can be scaled-up or down as per requirement, and has specific binding.

The Bad

Sometimes Protein A/G reacts to secondary antibody to give a non-specific band, and elution of proteins from the beads is difficult.

The Bottom Line

A simple and useful reagent for day-to-day immunoprecipitation experiments.

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