Anti-CDKL5 Antibody

September 15, 2017

BlueskyReddit
Molecular Physiology and Biophysics
Baylor College of Medicine
Other

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Company:

Abcam

Product Name:

Anti-CDKL5 antibody

Catalog Number:

ab22453

We wanted to test this antibody for its ability to detect Cdkl5 protein in immunofluorescence or western blot applications. It is horribly nonspecific for immunofluorescence and western blot. However, it does allow you to detect a band that corresponds to Cdkl5 which is specifically expressed only in animals that possess a wildtype allele of Cdkl5. 

Experimental Design and Results Summary

Applications

Western Blot

Sample

Mouse brain (frontal cortex)

Primary Incubation

1:500 4C overnight in 5%BSA, 0.1% Tween-20, 1xPBS

Blocking Agent

5%BSA in 1x PBS

Secondary Incubation

1:10k Licor 800 conjucated anti-rabbit prepared in 5%BSA, 1xPBS

Tertiary Incubation

N/A

Detection

Licor imager

Results Summary

We observed that this antibody was fairly nonspecific and detected several other proteins that are not Cdkl5. However, you can distinguish a band in samples taken from wildtype animals that is diminished in Cdkl5 mutant animals. The Sigma antibody is a better antibody because there are fewer non-specific bands near the molecular weight of Cdkl5 so the background is not as much of an issue for quantification. This antibody works, but I hesitate to recommend it. Hopefully, better antibodies for Cdkl5 will become available.(Note - this antibody also performed poorly for immunofluorescence due to its lack of specificity.)

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

It does detect a band that is probably Cdkl5

The Bad

It also detects several other non-specific proteins

The Bottom Line

This antibody is an option if you need to detect Cdkl5. In our hands the Sigma antibody is better, but both still detect several non-specific proteins.

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