When I studied the expression of Hypoxia Inducible transcription factor 1a from the whole cell lysate in western blot analysis, I got its expression. But when I isolated the proteins from cells using a nuclear lysate protocol, then there was no expression of the protein.
I had induced hypoxia using cobalt chloride at 200uM concentration for 24hrs.
Eugen Werwein
I would guess that your cell fractionation protocol is not optimal. Do you have a positive control, like histone staining to confirm nuclear protein enrichment?
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Divya Vipin Menon
Thank you Eugen Werwein for your repsonse.
No, I did not have histone staining to confirm nuclear protien. I only had a beta actin.
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