We are trying to replicate published data (so far without success) and we want to test whether our dCas9 construct is expressed. Since Cas9 untagged, we are looking for a well functioning antibody.
Anti-CRISPR-Cas9 antibody - abcam.com
Adwww.abcam.com/CRISPR-Cas9-antibody
Excellent Cas9 antibody for WB, IHC tested and reviewed by customers.
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Yes, but the reviews are only by researchers that used the antibody in eukaryotic system. There is presumably (due to its bacterial origin) more background in bacterial systems. Hence my specific search for an antibody that works well to detect (low) Cas9 levels in E.coli.
We can't vouch for E. coli reactivity, but we do list some other anti-cas9 Abs, both monoclonal and polyclonal, which may be worth exploring. Just search here http://www.linscottsdirectory.com/search/antibodies for cas9, and follow the "More Info" links to the suppliers' data pages.
If you still cannot find e.coli specific Cas9 antibodies, we do offer a monoclonal anti-Cas9 in a small volume that is low priced: http://www.epigentek.com/catalog/crisprcas9-monoclonal-antibody-7a9-p-4886.html
We also offer a polyclonal pAb if that may be of interest, but probably will not work well for e.coli as it has very high affinity and may result in some background. http://www.epigentek.com/catalog/crisprcas9-polyclonal-antibody-p-4812.html
Good luck!
I hope you may have found an antibody which works well in your experiments. However, for predicting the cross-reactivity, it would be helpful to have the exact sequence of the protein which you have over-expressed. Novus antibodies for this target are generated against immunogen sequences from CRISPR-Cas9 of Streptococcus pyogenes (UniProtKB - Q99ZW2; CAS9_STRP1) and we can check for immunogen sequence homology with your overexpressed protein using BLAST analysis. At Novus, CRISPR-Cas9 antibody (clone 7A9-3A3) [NBP2-36440] is very popular and it is good for WB, ICC/IF and IP. Iit has been cited in publications also and is available in unconjugated as well as conjugated formats (15 Fluorochrome options including Alexa Fluor^R and DyLight conjugates) http://www.novusbio.com/CRISPR-Cas9-Antibody-7A9-3A3_NBP2-36440.html Other options include a newer mAb CRISPR-Cas9 antibody (clone 6G12) [NBP2-52398], and two rabbit polyclonals CRISPR-Cas9 Antibody [NBP2-52718] and CRISPR-Cas9 Antibody [NBP2-52717]. All of these antibodies are highly validated and the images are available on their respective datasheets.
Thanks. We used meanwhile successfully Epigenetics CRISPR/Cas9 Polyclonal Antibody A-1111.
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