The citrate buffer comes in a 100x and 10x option. The 10x version includes a pH of 6.0 and comes in 125 ml while the 100x version comes in a 50ml version with a pH of 6.0, as well. Both versions are intended to be used for research and shouldn’t be used for diagnostics.
The Description
Primarily, a citrate buffer should be diluted appropriately and used on paraffin-embedded or Formalin-fixed tissue sections. These sections should be mounted onto glass slides before IHC procedures. When you perform IHC procedures with this reagent and a higher temperature, retrieval may disrupt the covalent bonds that are formed by the formalin in the tissues. When you remove those bonds, you allow renaturation of the protein molecules and can increase accessibility to the antibody, which results in better antibody binding and deeper staining. Likewise, this option can improve your signal to noise ratio.
More Information
The citrate buffer works well for Immunohistochemistry applications and contains the product with a pH of 6.0, either in 10x or 100x concentrations.
The staining protocol is to rehydrate and deparaffinize the tissue sections and dilute the citrate buffer appropriately. For the 10x concentrate, you will dilute using a ratio of 1:10 with deionized water. For example, if you use 20 ml of the concentrate, you should use 180 ml of deionized water for dilution.
If using the 100x concentrate, you will need to dilute the product using a ratio of 1:100 and deionized water. For example, two ml of concentrate would require 198 ml of deionized water.
Next, you will immerse the sections in a Coplin jar filled with your diluted citrate buffer reagent and microwave on HIGH until it is boiling. The slides must be warmed on LOW for 10 minutes. The Coplin jar should sit inside the microwave for 20 minutes, allowing it to cool slightly. Remove it all and rinse.
The Citrate Buffer is an excellent product to be used for IHC applications. Visit Spring Bioscience to learn more and purchase some for your laboratory.