Antigen retrieval refers to any technique in which the masking of an epitope is reversed and epitope-antibody binding is restored. Why is antigen retrieval necessary in immunohistochemistry (IHC)? The impaired ability of antibodies to access epitope in fixed tissue can impact IHC staining. Protein fixation creates crosslinks that mask tissue antigen and limit antibody-epitope binding.
2 mg/ml pepsin in Tris-HCl pH 2.0
Or 0.5 mg/ml trypsin in Tris-HCl pH 8.0
1. Pre-warm the buffer to 37°C;
2. Keep the sections in retrieval buffer at 37°C for 10-20 minutes.
3. Suitable for antigens that are sheltered by fixative such as:
4. Collagen, GFAP, Complement, Cytokeratin, C-erB-2, LCA, LN, etc.
10 mM Sodium Citrate pH 6.0
1. Microwave the antigen retrieval buffer until boiling and power it off, put in the sections;
2. Repeat step 1 for one or two more times for every 5-10 minutes;
3. Cool down at room temperature and wash with dH2O.
Bcl-2, Bax, c-myc, E-cadherin, ChromograninA, Heatshock protein, HPV, P-glycoprotein, PKC, PCNA, etc.