can anyone please let me know the difference between bca and micro bca assay which is used in protein quantification. In literature they used micro bca assay to determine liposome associated proteins.
Thank you.
Hey Harshavardan
The two are essentially the same (reagent wise) except for the volumes and reagent to protein ratios used. Have a look at the manual below, "test tube" vs. "microplate assay".
You can basically adjust the format to your specific needs using any intermediate setup as well...
Both Micro BCA and Pierce BCA assays can quantify protein via either 'microplate' or 'test tube' procedure. Microplate procedure in general requires less standard/sample volume. It depends on what protein concentration range your samples are within.
Pierce BCA microplate working range is from 20-2000 ug/ml, and it employs adding sample:reagent in 1:8 ratio - there may be some reagent interference and it may be hard to precisely quantify protein concentrations at the lower end of working range. The upside is that you only need to incubate it for 30 min.
Micro BCA microplate working range is from 2 - 40 ug/ml. You add sample:reagent in 1:1 ratio which is more desirable as opposed to Pierce BCA assay. You do have to incubate it for 2 hrs.
In regards to test tube procedures, they require larger sample volumes (1 ml of standard/sample/reagent for Micro BCA, or 0.1 ml standard/sample and 2 ml reagent for Pierce BCA). Further, all samples have to be analyzed via spectrophotometer within 10 min upon end of incubation, which may be less feasible when working with tens/hundreds of samples.
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