Western Blotting detection Antibodies........... - Are anti-sheep and anti-goat HRP interchangable? (Jul/20/2006 )
My colleagues here at work have almost convinced me that detection of primary antibodies from Sheep on a Western blot can achieved with both a anti-sheep or a anti-goat HRP secondary antibody.
Does anyone else have any experience of this.
Any opinions at all are greatly appreciated.
I have used goat secondary for sheep primary. I cant say I had huge success as that particular primary antibody was dirty.
However on another secondary antibody we used I noticed it said "Anti-Sheep/Goat". I never bothered to find out what animal it was raised in but I doubt that they would mix two solutions of secondary sheep & goat! It worked a treat too.
seems a anti-sheep/goat is an anti-sheep which has been selected for affinity to goat:
"The pooled antiserum is used to produce an immunoglobulin preparation which is then purified on affinity columns of sheep Ig. After washing to remove any remaining non-specific serum components and low affinity antibodies, the sheep Ig specific antibodies are eluted using
carefully selected, mild conditions which minimize aggregation and preserve immunological activity, yet which elute high affinity antibodies. In addition, the anti-sheep Ig antibody has been selected for high binding to goat Ig."