[Histonet] controls and secondaries

[Till, Renee]

What is the difference between  a secondary such as a goat anti- mouse
and a straight mouse IgG istotype control as far as it's applications in
the IHC process? I ask because someone from another lab recently came to
me for help with a long list of stains they need done in a short period
of time. They have already ordered everything they think they need to
complete the stains. Most of them are flouescent conjugated primaries,
but two will have to have the fluorescence attached to the secondary. I
am wondering about this because I was asked why their protocol would
call specifically for goat serum to block, and would it matter if it was
another species. I told them that usually the blocking serum is the same
as the species the secondary was made in and was told that the secondary
wasn't made in anything different such as a goat anti-mouse, they had
what I would normally use as control serum. Can it be used as a
secondary? I have never tried this. Supposedly it worked with their flow
samples as a secondary (these are mouse primaries on pig tissue).

 

Renee' Till, HT

 

Arkansas Childrens Nutrition Center

1120 Marshall

Little Rock, AR 

72202

 

(501) 364-2774