How To Use A Citrate Buffer

by | Sep 1, 2017 | Healthcare

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The Citrate Buffer can be utilized for research and should be diluted appropriately. It is used on formalin-fixed and paraffin-embedded tissue sections that are mounted on glass slides to target the retrieval of the antigen in Immunohistochemistry procedures. When you use this reagent and an elevated temperature, the target retrieval may disrupt covalent bonds formed by the formalin in the tissue. Removal of the bonds allows for renaturation of the protein molecules and an increase in antibody accessibility, which means an increase in antibody binding, and better staining intensity.

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You can find kits that include a 50ml size of 100x Citrate Buffer, as well as 125ml with 10x and a pH of 6.0.

Staining Protocol For 10x Version

Deparaffinized slides must always be used, and you should rehydrate the tissue section. You’ll need to dilute the Citrate Buffer using a ratio of 1:10 and deionized water. For example, if you must use 20 ml of the concentrate, you would use 180 ml of deionized water. Next, you’ll need to immerse the section in a Coplin jar filled with the diluted reagent, microwaving on high power until it starts to boil. The slides must be kept warm and heated for 10 minutes using low power. Afterward, you’ll let the Coplin jar sit for 20 minutes in the microwave. You can then remove it and rinse it with water. You should then rinse with the buffer and use your staining protocol for the antibody you’re using.

Staining Protocol For 100x Version

For the 100x version, the protocol is the same, though you will need to dilute the product using a ratio of 1:100 and use deionized water, as well.

The Citrate Buffer is used for staining purposes in IHC applications. So that you never run out, visit Spring BioScience for purchase options today.

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