Endogenous Biotin Interference (Immunohistochemistry (CST Guide))

Question

Accepted Answer

High background staining specifically in kidney, liver, or other biotin-rich tissues when using biotin-based detection. Background not resolved by standard blocking procedures. Common causes: 1) Biotin-based detection system used on tissues with high endogenous biotin levels (kidney, liver) 2) Endogenous biotin binds avidin/streptavidin detection reagents creating false-positive signal 3) No biotin blocking step performed prior to primary antibody incubation Solutions: 1) Switch to polymer-based detection system (SignalStain Boost IHC Detection Reagents #8114 or #8125) which eliminates biotin interference 2) If biotin-based detection must be used, perform biotin block after normal blocking procedure and prior to primary antibody incubation 3) Avoid biotin-based systems entirely for kidney and liver tissue IHC experiments