Fig. 4: Antibody validation by ELISA. | Communications Biology

Fig. 4: Antibody validation by ELISA.

From: Epitope-directed monoclonal antibody production using a mixed antigen cocktail facilitates antibody characterization and validation

Fig. 4

a Antibody-pair dose response against recombinant hANKRD1 (OriGene Technologies). The legend lists each capture/detection antibody matched pair used. A linear curve fit over analyte concentration ranging from 0.313–20 ng/ml is applied to representative antibody pairs. b Dilution curves (fitted with power-law regression where the exponent of function x represents the slope of the curve) of endogenous rANKRD1 and ectopic Myc-hANKRD1 present in transfected H9c2 lysate and hANKRD1 from induced E. coli cells. Data points represent 10, 20, 40 and 80 fold dilution of cell lysates with starting total protein concentration at ~2 mg/ml. Both axes are plotted in the log10 scale for better visualization of parallelism. c Effect of peptide competition in ELISA where the detection antibody is blocked by matched or unmatched peptides. Data from 10, 20, 40 and 80 fold diluted lysates are shown. d Plasma rANKRD1 concentration (ng/ml) in experimental MI and Sham rat on days 7 (D7) and 30 (D30). The boxplot represents data quartiles (25th, median and 75th percentile) and error bars represent the maximum and minimum data values. Data distribution for each boxplot is indicated by the coloured dots. Two-tailed unpaired t-test was used to compare each two-group combination with significance at p < 0.05. n number of individual animals in each group.

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