Immunohistochemistry Staining SOP
Purpose
To outline the standardized procedure for performing immunohistochemistry staining to ensure consistent and reliable results.
Scope
This SOP applies to all personnel performing IHC staining in the laboratory.
Responsibilities
- Technician: Perform staining according to SOP
- Quality Manager: Ensure compliance with SOP
- Lab Supervisor: Oversee training and adherence to protocol
Materials & equipment
- Formalin-fixed paraffin-embedded tissue sections
- Primary antibodies
- Secondary antibodies
- Blocking solution
- DAB substrate
- Mounting medium
- Microscope slides
Procedure
- Prepare tissue sections on microscope slides by deparaffinizing and rehydrating them.
- Perform antigen retrieval as per antibody requirements.
- Block non-specific binding sites using blocking solution.
- Dilute primary antibody in appropriate buffer and apply to sections.
- Incubate slides with primary antibody for specified time and temperature.
- Wash slides with buffer to remove unbound primary antibody.
- Apply secondary antibody and incubate as per manufacturer instructions.
- Wash slides again with buffer.
- Develop slides using DAB substrate and monitor under microscope.
- Counterstain if necessary and mount with appropriate medium.
- Allow slides to dry completely before storage.
Records
- IHC staining log
- Antibody lot tracking log
- Quality control records
Safety
- Wear appropriate personal protective equipment (PPE) such as gloves and lab coat.
- Handle all reagents according to their safety data sheets (SDS).
- Dispose of hazardous waste according to laboratory protocols.
Disclaimer
This is a template for educational and planning purposes. Laboratories must adapt it to their institutional SOPs, manufacturer instructions, applicable accreditation requirements, and pathologist approval before use.
Unimeditrek Pvt. Ltd. · www.unilabequipments.com · Educational template — adapt to your laboratory.