The product Citrate Buffer (pH 6.0, 10x) contains a 10× concentrated formulation of citrate buffer (pH 6.0) that can be used in Heat Induced Epitope Retrieval (HIER) methods. HIER is used to reverse the loss of antigenicity that occurs with some epitopes in formalin-fixed paraffin embedded tissues. During epitope retrieval, tissue slides are immersed in a solution and heated (>95° C) by hot plate or other appropriate heat source such as a pressure cooker or steamer. Citrate buffer is the preferred solution for most antibodies.
Antigen retrieval buffer for immunostaining.
Citrate Buffer (10×), pH 6.0 1L
Store at room temperature.
- Deparaffinize sections in 2 changes of xylene, 5 minutes each.
- Hydrate in 2 changes of 100% ethanol for 2 minutes each, 95% and 75% ethanol for 2 minute each. Then rinse in distilled water.
- The Citrate Buffer (10x) should be diluted 1:10 for working solution (1x).
- Pre-heat steamer or water bath with staining dish containing 1x Citrate Buffer until temperature reaches 95-100 °C.
- Immerse slides in the staining dish. Place the lid loosely on the staining dish and incubate for 15 minutes (optimal incubation time should be determined by user).
- Turn off steamer or water bath and remove the staining dish to room temperature and allow the slides to cool for 20-40 minutes.
- Rinse sections in PBS Tween 20 for 2×2 min.
- Proceed to standard immunohistochemistry protocol.
Note: Microwave, pressure cooker and autoclave can be used as alternative heating source to replace steamer or water bath.
Note: This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals. Avoid contact with eyes, skin and clothing. Do not ingest. Wear gloves.
User Manual (PDF)