Suboptimal Primary Antibody Incubation (Immunohistochemistry (CST Guide))

Question

Accepted Answer

Inconsistent or weak staining results despite following protocol. Staining intensity varies between experiments using same antibody lot and tissue type. Common causes: 1) Primary antibody incubation time shorter than validated protocol (CST antibodies validated for overnight at 4°C) 2) Incubation temperature not controlled or inconsistent 3) Deviation from rigorously tested, product-specific protocol Solutions: 1) Follow CST validated protocol: incubate primary antibody overnight at 4°C for optimal, consistent results 2) Maintain consistent incubation temperature (4°C) across all experiments 3) Refer to product-specific datasheet for rigorously tested incubation conditions; do not modify without re-validation