This is my first time doing a BCA assay for my protein sample, So my question is should I make a serial different dilutions for my sample or make dilutions with the same concentration?
as the protocol suggested to add 5ul of the sample with 20ul of diluent for all samples.
My second question is for working reagent I found an example to calculate how much need for WR preparing, but I didn't understand it is supposed to be used 14.4 instead of 20.4 right?
could anyone help me please, Tq
Example:
8 standards × 3 replicates = 24 wells
3 unknowns × 8 dilutions × 2 replicates = 48 wells
(48 + 24) × 0.2 mL = 14.4 mL of RAB
Prepare the volume of required RAB by using the ratio of 50:1
Example (if 14.4 mL of RAB is needed):
(20.4 mL / 51 parts total) × 50 parts A = 20 mL Reagent A
(20.4 mL / 51 parts total) × 1 part B = 0.4 mL Reagent B
sample concentrations can be estimated using BCA using Standard curve , samples should be approximately diluted so that the measured concentration falls within the standard curve .
Kandluri Geethika Thank you for your response, I appreciate it
so should I make different dilutions for my sample like 1:2,1:4@ etc?
or make one dilution only for example 1:2 dilution for all selected wells
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