Monthly Archives: October 2016

How to calculated B/B0, % bound, for your competitive ELISA

Competitive ELISA is one of the many types of ELISA protocol. Competitive ELISA differs from sandwich ELISA in that the higher the concentration of the antigen in the sample, the lower the intensity reading. The maximum intensity is achieved when no antigen is present in the sample. To assess this maximum level in a competitive ELISA there is usually a control well in the plate layout that specifically contains no antigen and this is known as a Zero well or B0. Then the sample intensities can be compared to the B0 well to obtain a percentage bound. This is symbolized as B/B0 where B is the intensity of a sample well and B0 is the maximum intensity.

To calculate B/B0 with ELISAAnalysis.com your should specify the B0 wells in your plate layout in step 2 of the work flow and select the check box “Calculate % bound (B/B0) for all samples using Maximum Binding (B0) controls?”  in step 3. ELISAAnalysis.com will do the rest.

Adjusting to allow for Non-Specific Background (NSB)

ELISA is a robust assay, but it is inevitable that there will be some binding of proteins to the plate that is not intended. Using NSB wells in your plate layout will give you the option to subtract out this background noise from your intensity readings before analysis.

To do this with ELISAAnalysis.com your should specify the NSB wells in your plate layout in step 2 of the work flow and select the check box “Subtract Non-Specific Binding (NSB) Control well average OD from all other ODs?” in step 3. ELISAAnalysis.com will do the rest.

Data Transformations using control wells, B/B0, Zero wells, NSB wells

ELISA Analysis now has the ability to adjust your data (transform it) using the control wells in your plate layout before proceeding to the curve fitting and calculation steps.

Adjusting to allow for Non-Specific Background (NSB)

ELISA is a robust assay, but it is inevitable that there will be some binding of proteins to the plate that is not intended. Using NSB wells in your plate layout will give you the option to subtract out this background noise from your intensity readings before analysis.

To do this with ELISAAnalysis.com your should specify the NSB wells in your plate layout in step 2 of the work flow and select the check box “Subtract Non-Specific Binding (NSB) Control well average OD from all other ODs?”  in step 3. ELISAAnalysis.com will do the rest.

How to calculated B/B0, % bound, for your competitive ELISA

Competitive ELISA is one of the many types of ELISA protocol. Competitive ELISA differs from sandwich ELISA in that the higher the concentration of the antigen in the sample, the lower the intensity reading. The maximum intensity is achieved when no antigen is present in the sample. To assess this maximum level in a competitive ELISA there is usually a control well in the plate layout that specifically contains no antigen and this is known as a Zero well or B0. Then the sample intensities can be compared to the B0 well to obtain a percentage bound. This is symbolized as B/B0 where B is the intensity of a sample well and B0 is the maximum intensity.

To calculate B/B0 with ELISAAnalysis.com your should specify the B0 wells in your plate layout in step 2 of the work flow and select the check box “Calculate % bound (B/B0) for all samples using Maximum Binding (B0) controls?”  in step 3. ELISAAnalysis.com will do the rest.