I want to do antigen retrieval for my frozen section following by immunofluorescent staining. I am wondering what the difference between sodium citrate buffer vs. citrate buffer in terms of antigen retrieval?
Here are the recipes from IHC world.
Sodium Citrate Buffer (10mM Sodium Citrate, 0.05% Tween 20, pH 6.0):
Tri-sodium citrate (dihydrate) - 2.94 g
Distilled water - 1000 ml
Mix to dissolve. Adjust pH to 6.0 with 1N HCl and then add 0.5 ml of Tween 20 and mix well. Store this solution at room temperature for 3 months or at 4 C for longer storage.
Note: this buffer is commonly used and works perfectly with many antibodies. It gives very nice intense staining with very low background.
Citrate Buffer (10mM Citric Acid, 0.05% Tween 20, pH 6.0):
Citric acid (anhydrous) 1.92 g
Distilled water -1000 ml
Mix to dissolve. Adjust pH to 6.0 with 1N NaOH and then add 0.5 ml of Tween 20 and mix well. Store this solution at room temperature for 3 months or at 4 C for longer storage.
In the sodium citrate buffer you will have a small amount of sodium chloride which gives slightly higher ionic strength. I suspect that for your application either buffer is fine.
As far as I can tell, these buffers are the same. The difference is in how they are prepared.
Dear
According to me the difference between these buffer is due the presence of soduim in sodium citrate buffer. So sodium citrate buffer is commonly used and works perfectly with many antibodies. Also It gives very nice intense staining with very low background. Therefore I recommend you sodium citrate buffer
Regards
Marius
I agree with Adam. They are both sodium citrate, at least if you use NaOH to bring the citric acid up to pH 6. And, if you rely on citric acid itself as the H+ donor, in a 4.4:1 ratio of isosmolar solutions of sodium citrate:citric acid, you'll get to pH 6.0 without the need for HCl. See this hyperlink for a recipe for 0.05M citrate buffer by this method: http://microscopy.berkeley.edu/Resources/instruction/buffers.html
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