Question (75683) | Anti-STAT2 (phospho Y690) antibody (ab53132)

Go to datasheet (ab53132)

Question

Is the blocking peptide available? I am obtaining two bands at 100 and 115 kDa in my human cancer cell line.

Answer

Unfortunately, the blocking peptide for ab53132 is not available. In order to assist in your determing what the identity of your two bands are, I did some research and found that IFN-alpha and IFN-beta induce activation of Jak kinases, which lead to phosphorylation of STAT1 and STAT2. I found a nuclear lysate (STAT2 is translocated to the nucleus upon phosphorylation/activation by IFN alpha) in our catalog of U937 cells that were treated with IFN alpha. The catalog number of this antibody is ab14907:

https://www.abcam.com/u937-human-leukemic-monocyte-lymphoma-cell-line-nuclear-lysate-ifn-alpha-treated-ab14907.html

Alternatively, you can treat your own cells with IFN alpha as a positive control, and perhaps perform a nuclear fractionation of your samples to determine the identity of the two bands since phosphorylated STAT2 should be present in the nucleus. There is also the possibility that you are detecting both isoforms and they are somehow separating more than expected in the gel. A link to the uniprot file for STAT 2 depicting both isoforms is given below:

http://www.uniprot.org/uniprot/P52630

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