The R2 value is an indication of how closely the data fit the trendline. y=mx + b m= slope of the line and b= y intercept) and the R2 value. Use the computer software to generate the equation of the line (i.e. Typically, a standard curve will have a sigmoidal shape in which the higher concentrations of standard dilutions will reach a plateau in absorbance.įind the portion of the curve that is linear and draw the best-fit trendine for the data (many ELISA plate readers have built-in programs for generating and analyzing standard curves or you can use Microsoft Excel, or similar graphing software). Then generate a standard curve by graphing the mean absorbance for each sample (x-axis) vs. Once the intensity of each well has been measured on the plate reader, calculate the average absorbance values for each duplicate/triplicate sample. This is a brief guide to preparing a standard curve and using the standard curve to quantify protein amounts. The preparation of the ELISA standard curve is critical in accurate sample quantification. The amount of target protein within a sample can be precisely determined by setting up a standard curve of known target protein concentrations using purified antigen. Samples can be compared to one another to determine the relative amount of protein within each sample, as the intensity of signal is proportional to the concentration of the target protein. An ELISA can be used to determine whether the protein of interest is found within a particular sample by comparing the sample to a blank well or a sample that does not contain the target protein. When performing ELISAs, three different data outputs can be generated: The enzyme-linked immunosorbent assay (ELISA) is a sensitive technique for determining the concentration of a protein in a liquid sample.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |