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. Author manuscript; available in PMC: 2015 Dec 1.
Published in final edited form as: J Microbiol Methods. 2014 Sep 16;107:58–62. doi: 10.1016/j.mimet.2014.08.026

Fig. 2.

Fig. 2

Specificity of the ELISA assay. (A) CabP binds c-di-AMP specifically. Using coating concentrations of 10 and 50 μg/ml, CabP and BSA were coated into each well similarly. Subsequently, a titration of B-c-di-AMP ranging from 0 nM to 1,000 nM was performed in order to optimize efficiency and lower overall use of B-c-di-AMP. Our results from this titration indicate that a concentration of >1.5 nM was required for proper detected. (B) c-di-AMP binding site in CabP is specific for c-di-AMP. Using a 10 μg/ml coat of CabP and a standard 25 nM B-cdi-AMP tracer, 200 nM of various closely related nucleotides were used to challenge the performance of this assay. The only nucleotide that significantly reduced the signal was c-di-AMP, indicating that this assay is specific for c-di-AMP and not the other nucleotides tested. Data shown are the means of two independent experiments. The error bars denote the standard deviation (SD). Data set labeled “Control” represents repeats without any nucleotide challenge.