I am trying to using the Gen5 Cytation by BioTek for protein quantification using a micro BCA Assay kit.
I downloaded a BCA Assay Experiment (.xpt) File, and tried to use it but ran into some trouble. When setting up/entering info for the plate map, I was able to add the BSA standard wells with no problem because I knew the concentrations of each standard in ug/mL.
For unknowns it didn't let me specify my own dilutions. There were some options given (1:1, 1:2, up to 1:96) but it wouldn't let me choose the dilutions that I had since my dilutions didn't start at 1:1 and mine increased in larger increments.
Am I stuck using those dilutions suggested by Gen5 or does someone know a way to set up the protocol differently?
Hi Casey Hughes ,
Can you share your protocol? do you have varied dilutions of your unknown samples?
Yes, this is the file I found online. My dilutions of the unknown samples varied from 1:20, 1:40, 1:60, and 1:120. Thanks!
Unfortunately, I couldn't open the file. However, you can't use different dilutions of the unknown samples. For example, let us suppose you have three samples of interest (Control, factor 1 of interest-treated cells, factor 2 of interest-treated cells). Here you have to add the same amount of your samples (for example: 20 micro liter), dilute them at same rate with the working reagent (for example: 1:10) and measure the absorption. After that, you will draw a blot chart in comparison with the standard one to estimate the concentration of protein in your sample.
Hopefully this gives you a hand.
Best.....
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