I'm going to use BSA as standard for Bradford test. However, BSA is hard to dissolve. I searched around and there are different suggestions on how to dissolve BSA.
Some suggest layering BSA in solution or water. Some use PBS to dissolve BSA while some suggest to use protein extraction buffer (the same buffer that was used to extract protein from sample) to dissolve BSA.
Which is the best way to dissolve and prepare 1mg/ml BSA for standard curve in Bradford test?
BSA readily dissolve in warm distilled water or a buffer. Warm either of these to 38-39 C and add required amount of BSA to prepare a stock solution for use in preparation of the reference curve by Bradford assay.
1 mg/mL can be easily dissolved in plain water or a regular buffer, it just takes time at room temperature
BSA can readily be soluble in water at room temperature. However, be careful not to overmix it because it will cause bubble formation.
In my scenario, I take BSA (1mg/mL) and take buffer as a diluent for preparing the standard. Diluent must be the buffer used for making the lysate. If you are doing cell culture lysate's protein estimation, then you should use lysate buffer or RIPA (whatever you have taken for making lysate).
In my case, to prepare BSA standard curve, first I dissolve 0.1g BSA crystals in 10mL milliQ water to make a stock solution of 10 mg/mL at room temperature
From here I dilute in the range of 12.5-200ug/ml. I hope this is helpful for you.
follow the below link
https://assets.fishersci.com/TFS-Assets/LSG/Application-Notes/TR0006-Extinction-coefficients.pdf
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