Re: Double labeling on mouse
Hi, I think Gayle had given a good explanation in the past.
You basically do two reactions sequuentially.
The first detection is with Antibody A all the way till the DAB step.
Stop, place slides in a buffer (we use an Antigen retreival buffer,
Retrievit-8) and microwave for about 10seconds/slide. Wash well with
Wash with PBS.
Start the second detection redoing all the protein blocks (avidin-biotin
blocking if you are using that) all the way to the end with Antibody B.
Counterstain and mount.
We prefer the following chromogen pairs
DAB for the first reaction followed by AEC. Counterstain Hematoxylin.
BCIP/NBT for the first reaction followed by Fast Red.
Karen Pawlowski wrote:
> Hi everyone,
> I was wondering, has anyone done double labeling with a mouse on mouse
> kit? Would you need to have two different secondary antibodies?
> Karen Pawlowski, Ph.D.
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