Question (5523) | Anti-6X His tag® antibody (FITC) (ab1270)

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Question

I was wondering about the fixatives used for this antibody. Is paraformaldehyde ok, or must I use methanol, etc. More importantly, can this antibody be used on unfixed cells to detect the presence of a cell-surface HIS-tagged protein in unpermeabilized cells ?

Answer

The method of fixation is unlikely to have significant effect on the His-tag. However, the fixation may influence the conformation of the tagged protein, and thereby, affect the accessibility of the His-tag. Thus, one must empirically determine the influence of fixation upon ability to detect tagged proteins.

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