Which band do you look at (100 kDa, 68 kDa)?
Hi,
We are currently using Santa Cruz anti-NRF-2 antibody to detect NRF-2 expression. We see bands usually at 65, 68 kDa. However, sometimes a band around 100kda appears. Which one (s) are correct? Do they represent different states (degraded vs active)? If anyones knows of a better antibody and which band we should consider active, please let me know.
thanks in advance!
67.8 is the calculated MW for human Nrf-2.
http://www.uniprot.org/uniprot/Q16236
A shift to that range (approx. 100 kDa) can occur by N-linked glycosylation. To confirm this use PNGase in samples and see if that 100 kDa form disappears and 67.8 kDa form increases.
Kit: GlycoProfile™ II Enzymatic In-solution N-Deglycosylation Kit - See more at: http://www.sigmaaldrich.com/technical-documents/articles/biology/glycobiology/deglycosylation-kits.html#sthash.ldfNLpMa.dpuf
Our EB11042 detects a 70kDa in human lung lysates (see http://everestbiotech.com/product/goat-anti-nrf2-aa445-458-antibody/). However, we do see a ~100kDa band in some rodent tissues with this antibody. It might be worth comparing side-by-side.
Nrf2 antibody from Genetex works best for me. It is detected approximately 92 kDa. For more Details regarding its molecular weight , this link might be helpful http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3503463/
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