Hello! I am using an anti-NFATc1 Santa Cruz antibody (7A6, sc-7294). The company predicts the molecular weight for the isoforms in 90; 110 and 140 kDa. My problem is that I hardly get to see these bands, and I have a very strong band between 37 and 50 kDa.
On the other hand, Sigma for its anti-NFATc1 shows a band between 40 and 65 kDa, although it predicts that the band is at 78 kDa.
Did something similar happen to someone?
hi
about 103 kD
probably your samples express one isoforme (NFATC1-204) from 10 transcript . To further ensure the antibody's performance, you can use Raji & Jurkat cell lysate along with your samples and put your test in non-reducing condition .
https://www.rndsystems.com/products/human-nfatc1-antibody_af5640#ds_image_0
http://www.ensembl.org/Homo_sapiens/Gene/Summary?db=core;g=ENSG00000131196;r=18:79395856-79529325
good luck
thanks for helping!
The gel is 10% polyacrylamide and cells are BMDM.
Dear Ximena Volpini
You have got a band with the same intensity in all your samples. it is more likely that your Ab has a strong non-specific reaction with a house keeping gene product. 10% gel is OK. Use a positive control cell line. if you dis not your desired band change your Ab. Some Santa Cruz Abs are not good. I have used NFATc Ab from R&D. It is very good.
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