Hi, good day...I conducted a human study toward the end of 2019, and I have kept my samples at -80 degrees since. Now I would like to analyse the expression of transcription factors (Nf - kB and Nrf2, Western blot analysis). I have whole blood and buffy coat samples aliquots that I thought I could use to isolate PBMCs but my worries now is that these samples have been stored at -80 degree for that long, and all the PBMCs isolation protocols I have seen so far it seem only required fresh blood samples. .

If anyone has encountered or experienced or know of a protocol I can use or follow to isolate PBMCs from a buffy coat or whole blood sample pls help on the possible protocol/ procedure?

Thankx very much.

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