Biotin contamination of BSA
Dr Engelbert Buxbaum
via methods%40net.bio.net
(by engelbert_buxbaum from hotmail.com)
Thu Jul 16 10:50:59 EST 2009
Am 16.07.2009, 05:54 Uhr, schrieb WS <novalidaddress from nurfuerspam.de>:
> I noticed that "ordinary" BSA is unsuitable for blocking westerns when
> streptavidin HRP is used, as it causes high background by its biotin
> content. Where does the Biotin contamination come from and how may I
> get rid of it (except buying biotin free BSA)? Is it covalently
> linked?
Try fish skin (teleost) gelatin instead, or polyvinylpyrollidone (PVP). If
you blot on PVDF membranes, you do not need to block at all. Air-dry the
membrane completely, then all solutions can interact only with
surface-bound proteins, but not with the very hydrophobic membrane -> no
background binding.
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