how to block endogenous biotin?

Henk Veldman H.Veldman at lab.azu.nl
Thu Oct 14 02:18:47 EST 1999



dcoovert schreef:

> I need to block endogenous biotin in a tissue section. I have the
> reagents: avidin, d-biotin. I however do not have a protocol. (My
> protocol book recently left with a former student -AND IF YOU ARE HE...
> BRING IT BACK!) Thanks in advance for any help. Thanks
>                         -Dan

1 - Rinse in PBS . . . 10 min
2 - 0,01 % - 0,1 % avidin in PBS . . .  20 min
3 - Rinse in PBS . . .  5 min
4 - 0,001 % - 0,01 % biotin in PBS . . . 20 min
5 - Rinse in PBS 5 . . .  min
6 - Proceed with immunohistochemical procedure . . .

When making stock solutions, remember that biotin does not dissolve very
well in water: at most about 20 mg in 100 ml at room temperature, less so
in the fridge.

Reference: Woods & Warnke; J Histochem Cytochem 29: 1196-1204, 1981.

Success

Henk






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