Determination of Sample Concentration

[Guest NNN]

Dear Mabtech support Team,

I have a question regarding the determination of the sample preparation. So for normalizing the standard curve, I subtract the Blank from all values; but for my samples I also have Blank wells (control/only medium e.g.) measured. So should I also subtract the Blank from my sample values, before using the value for the equation or not?

Thanks a lot for your help!

Best, NN 

[Christian@mabtech.com]

Hi NN,

 

At Mabtech when we say blank we refer to the the "plate blank" - that is when only substrate and stop solution is added to the wells. This is subtracted by the software before creating the standard curves.

 

We know that not all customers use this kind of blank, but instead chooses to subtract the control with only buffer (incubation buffer or ELISA diluten). Your question, however, concerns whether we recommend subtracting the background values obtained from samples with only medium, e.g. unstimulated control cells. These we do not recommend to subtract, so the short answer to the question would be - no.

 

The wells without analyte, buffer only, mock, control etc. is preferably shown. These values will give an indication of the quality the assay in terms of background.

 

 

[Guest NNN]

Hi Christian,

thanks a lot for coming back to me! Yes, for creating the standard curve, I subtract the 'plate blank'. I was just confused because I've got a recommendation from a colleague to also subtract the 'blank (=medium only)' from the measured samples before calculating. But as you said it makes sense to not do that, just wanted to ask experts:) Thanks again for your help!

Best, NNN