Protocol Online logo
Top : Forum Archives: : Immunology and Histology

Immunohistochemistry blocking - Whether to use or not? (Jul/30/2008 )


I am doing some immunohistochemistry using the LSAB kit from DAKO. The instructions for this kit state that a non-specific blocking step is not required. I have previously always used a blocking solution of some sort. I have performed a few runs without it though and have found that I get good, seemingly specific staining.

Ive also tried a few different blocking buffer additions to the protocol (BSA, Milk, plus a few commercial varieties) and it doesn't seem to improve the results.

So basically its easier to do the runs without the addition step, so I was wondering if there were any opinions on publshing IHC results that did not use a blocking step.

As I said earlier I have always used a blocking step previously and want to get peoples ideas before going ahead with my samples.



I would say if it works and you don't get non-specific staining and you can't improve I see no reason to do the block.
Publishing results is for what works for you and if you really want to add it you can put a special note in your material and methods saying you've tried w/ and w/o block and got the same result.