Cell Fractionation Kit Review

January 31, 2017

Aquinnah Pharmaceuticals
Research
Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

Abcam

Product Name:

Cell Fractionation Kit

Catalog Number:

ab109719

We bought this kit thinking it would be an easy way to run a subcellular fractionation, to repeat data from a paper that we saw. We followed the instructions and used the same antibodies as the product insert for controls. The controls appeared across multiple fractions meaning that the fractionation was dirty if it worked at all.

Experimental Design and Results Summary

Application

Subcellular Fractionation

Starting Material

PC12 cells

Protocol Overview

Spin down cells and resuspend pellet in Buffer A, add Buffer B and incubate. Centrifuge and separate pellet and supernatant. Respin supernatant to get final C supernatant. Resuspend pellet in Buffer A and add Buffer C, and incubate. Centrifuge and separate pellet and supernatant. Respin supernatant to get final M supernatant. Resuspend pellet in Buffer A for N fraction

Tips

Did not work for us.

Results Summary

The three fractions were not pure and our control signal was found throughout the fractions.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Theoretically easy to do and doesn't involve prior cell lysis

The Bad

Did not produce clean separation

The Bottom Line

Although seemingly easy. Following the protocol did not yield clean fractions, with the control markers in multiple fractions for each. We made our own buffers and did the classic protocol and it worked great. Don't bother with this kit.

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