Dear all, I would like to optimise the concentration (%) of BSA to use for diluting my primary and secondary antibodies (in 1x PBS) to be used for immunofluorescence staining (and subsequently confocal imaging). I am curious to know what is the best concentration of BSA do you guys use? For your kind advice, please. Thank you :))))
Dear Justin,
We always use 1% BSA in Phosphate Buffered Saline. Never tested any other concentrations because results were good.
With kind regards,
Ronald
Hi Justin, I am working with 1% BSA in PBS/TBS with 0.2% Triton-X100 or 0.05% Tween 20 for Immunohistochemistry as well as for Insitu hybridisation with good results. If you are using lyophilisated BSA I recommand to filter the solution befor use to prevent that small unsoluble cristals are getting onto the sections.
You do not always need BSA as a "block" in IHC reagents. If you do then use a quality BSA for IHC staining. 1%-5% in PBS or TBS are commonly used concentrations, depending on your primary and your tissue sections.
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