I am staining for intracellular pSTAT1 and I am repeatedly running into difficulty owing to the fact I have to permeabilise with methanol (for sufficient nuclear entry of anti-pSTAT1 antibody and to facilitate STAT dimer separation).
The methanol is wreaking havoc with certain fluorochromes/antigens and causing either no successful staining, or false/bizarre stains. The problem is that it is proving almost random as to which fluorochromes/antigens are sensitive and which are resistant, for example a certain fluorochrome will work for one particular antigen and not another, and vice versa, suggesting it's a combination of both.
As of yet, I haven't found a reliable source of information to guide the design of my experiments around this, and was wondering if anyone had any ideas or know of any resource that may help me!
Thanks in advance!
http://media.ebioscience.com/data/pdf/best-protocols/staining-intracellular-antigens-for-flow-cytometry.pdf
On page 8 of this document, it lists some of the methanol-resistant and methanol-sensitive fluorochromes. It isn't too extensive, but it's helped me on more than one occasion.
Here is another good list:
http://www.bdbiosciences.com/documents/antibodies_human_cellsurface_marker.pdf
Unfortunately the new fluorochromes are not yet in the list.
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