I assume that you are using a biotinylated detection system? If so, endogenous biotin labelling is specific, tho' NOT wanted ;-) So, before you try variations, just buy a kit and see if your "non-specific" immunostaining is due to endog. biotin ...follow their instructions by pre-incubating in the kit. Then, if you find that the staining you get is due to endog. biotin...you can modify away ( such as including in primary Ab: However, the kits are RTU: so, by adding to primary Ab, for eg, you use a heck of a lot of kit solution.....;-)
I always pre-incubate because there are so many tissues that do NOT require endog. biotin blocking. Also, if you discover that endogenous biotin is the problem and you will therefore need to use a biotin blocking kit regularly, why not make up your own? Far cheaper/equally effective: many sites give protocols, such as this one http://www.immunoportal.com/ Please Post further if you have any problems. Good luck. _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
