We did the Pierce BCA assay and ran the standard curve (and were able to estimate protein concentrations in our samples). Is there a assay better than BCA for protein estimation? BCA assay - folded or denatured protein (Jul/15/2005 ) Hi all I run a BCA assay (bicinchoninic acid assay) to determine the concentration of protein in cell extracts.
The Pierce Modified Lowry Protein Assay is based on the colorimetric method introduced by Oliver H. Lowry in 1951. Protein measurement after Proteinase-K digestion? Does that mean the out put values are wrong and I cant trust them...?? t-Octylphenoxypolyethoxyethanol,
Why would 2D-GE analysis be too complicated? RIPA is just a mixture of detergents and buffering agents.
You may need a second elution step (with a volatile organic solvent) to remove the BSA. Accuracy of the Pierce Rapid Gold BCA Protein Assay and Bradford Protein Assay with known protein mixes. Product Results, Synonym: Can you share with me how much volume (ul) of unknown sample can be used for the dilution? Can streptavidin peroxidase (S5512) inhibit the results of a BCA Protein Assay? my Culture medium is BMM( (100 mM phosphate buffer(pH 6)/1.34% (w/v) yeast nitrogen base/1% (v/v) methanol/ 4 × 10−5% (w/v) of biotin). You can try to find a paper that reference the concentration of your particular specie or if you can't find it, do a serial dilution with a non-precious sample and see how much you have to dilute your sample so that they can fit on your curve. RIPA's advantage is that it's very denaturing while giving very low background when using copper based methods like BCA. We had excess standards so we decided to save them and are currently stored in -80 freezer. Hi allI run a BCA assay (bicinchoninic acid assay) to determine the concentration of protein in cell extracts. The things that can cause background on a BCA are thiol reducing agents, which will make it look like there is a higher protein concentration, and chelating agents, which will bind to copper (II) and prevent it from reacting. Our goal is to use colorimetric method (BCA assay) to quantify the protein content in these samples. Could the problem here be that only some of the protein is migrating through the gel? Why do we need add beta-mercaptoethanol in sample buffer to determine Bromelain's MW in SDS-PAGE? In this case we need to select a good detergent that doesn't interfere with the following precipitation step with TCA 50%/Acetone. The BCA protein assay is a simple and reliable protein quantification method. Is there any other method to quantify individual viral proteins apart from this? I used the same procedure on Jurkta cells. I Have two different protocols and one does and the other does not. are looking good. Both assays were conducted according to the manufacturer’s protocols, in a microplate format.
and since I loaded such a small amount of protein (1.95ug in an effort to save sample), I am not able to detect bands for these markers. Can you post a summary of it? Transfer success I have not checked with Ponceau staining yet [just the ladder transfer check], I have made some adjustments to the protocol but I am mostly attributing the issue to poor sample preparation.
In any case, the red color itself may not be a problem for the BCA assay, as long as it is not too much. its always better to use filtered sterile PBS.
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