Immunostaining of mouse spleen with rabbit antisera

Brian Devlin devlin at darwin.stanford.edu
Tue Jul 8 23:57:54 EST 1997


In article <K_Vora-3006971718400001 at manser3.jci.tju.edu>,
K_Vora at lac.jci.tju.edu (Kalpit A. Vora) wrote:

>Dear Readers,
>I am trying to stain mouse splenic cryosection with an polyclonal antisera
>raised in rabbits. I am having a terrible background staining problem. I
>do block with buffer containing 5% normal mouse serum(NMS) and 5% normal
>goat serum(NGS). Also all dilution of my primary and secondary antibodies
>are done in the above buffer containing NMS and NGS. Any pointers to
>improve my staining and decrease my background will be very helpful and
>thankyou in advance for the same. My e-mail address is as follows:
>kvora at lac.jci.tju.edu.
>Kalpit A Vora

It seems to me that blocking with pre-immune rabbit serum would make more
sense, but I don't actually have much experience with polyclonal sera.



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