Hello All,
I have been using the Albumin standard ampules from thermo scientific of 1ml to make a standard curve for protein and I even got a nice standard curve.
But when I am using the solid BSA flakes from Sigma Aldrich to prepare the solution required and when I am doing the Bradford assay with this sample, the colour is not changing or can say the Bradford reagent is not working on this solid BSA sample to detect the protein.
The ampules are of 2mg/ml in a 0.9% aqueous NaCl solution containing sodium azide. But the solid BSA flakes from Sigma Aldrich is a pure protein.
So, is this the reason for not detecting the protein through Bradford reagent on solid BSA flakes?
If anyone encountered the same issue please let me know.
And also, if some one has a solution to solve this problem, I would really appreciate it.
Thankyou,
Navneet Kallapalli
Crystalline BSA is a little difficult to dissolve in water because it is very hydrophobic. Nevertheless, once you have made the solution, it should work well for the Bradford assay standard. I prepare a 10 mg/mL standard. Start with a clean beaker. Add distilled water and a stir bar. On a magnetic stirrer, get the water stirring very rapidly, making a strong vortex. Gradually add the crystals of BSA and allow plenty of time for them to dissolve fully. You can store aliquots of the standard solution in the freezer so you don't have to make a new solution every time.
Thankyou Adam,
So it means it doesn't matter if you you crystallized BSA or albumin ampules for the standard, it will still work.
Because the ampules are having some chemicals mixed in the BSA and the crystallized dont.
The only chemicals in the ampules besides BSA are NaCl, which has no effect on the Bradford assay, and sodium azide as a preservative, which is also negligible. Both the BSA from ampules and crystalline BSA should give the same result, as long as the crystalline BSA is fully dissolved.
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