I’m working on NPAS2 in cancer. I’m trying WB. The predicted band is close to 100KDa. Sometimes I see a band around 75KDa.
Abcam’s antibody shows ~75KDa. https://www.abcam.com/npas2-antibody-ab137601.html
Do anyone have experience working with NPAS2 and know how to explain this? For example what sort of cleavage (if this is true) is happening, or is the antibody specific?
when I use a over-expression plasmid (NPAS2-FLaG) I do see correct ~100KDa band.
thanks!
Hey,
it is possible to be the work of proteases known as kallikreins (my thesis was on PFK1) and we also got shorter fragments (45-47kDa and 72 kDa instead of 85 kDa). They apparently tend to be overexpressed in cancer, but I'm not sure if it applies specifically to your case. You can check the article below:
Goettig, Peter, Viktor Magdolen, in Hans Brandstetter. 2010. „Natural and synthetic inhibitors of kallikrein-related peptidases (KLKs)“. Biochimie 92 (11): 1546–67
And I'm not saying it is the reason, but it is a possibility.
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