Re: [Histonet] BCIP/NBT with blocking buffer

From:Gayle Callis

The answer is theroretcially - yes.  However, this is not the endogenous 
enzyme you need to block for this chromogen.   NBT/BCIP is a chromogen for 
alkaline phosphatase method, so blocking endogenous peroxidase is 
unnecessary. You need to put levamisole in with chromogen to block 
endogenous alkaline phosphatase found in tissues.  Instruction sheets 
usually tell you this.  Levamisole is sold separately i.e. Vector has it.

  OR are you doing a double stain?  with both HRP and AP methods.  If that 
is the case, you would still need to block for endog perox for the the HRP 
portion of staining AND block with levamisole with AP portion.

At 12:18 PM 12/22/2004, you wrote:
>Hello. Can anyone tell me if I can block endogenous peroxidase with 0.3%
>H2O2 in sodium azide and still visualize with BCIP/NBT on frozen sections?
>Histonet mailing list

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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