CRM197 ELISA Assay Kit
The CRM197 ELISA Assay Kit Assay Kit is For Research Use Only
Size: 96 wells
Standard Range: 0 – 800 ug/mL
Sensitivity: 3 ug/mL
Incubation Time: 3 hours 30 minutes
Sample Type: Cell culture Supernatant, vaccines, and biological preparations
Sample Size: 100 µL
Assay Background
CRM197 is a genetically detoxified form of diphtheria toxin. A single mutation at position 52, substituting glutamic acid for glycine, causes the ADP-ribosyltransferase activity of the native toxin to be lost. CRM197 is widely used as a carrier protein for conjugate vaccines. A potential advantage of CRM197 over toxoided proteins is that, because it is genetically detoxified, it retains its full complement of lysine amines for conjugation.
Assay Principle
The method employs sandwich ELISA technique. Plates are coated with Streptavidin. Biotinylated Anti-CRM antibody is added to the plate and incubated. After incubation wells are washed with wash buffer and standard and sample are added to the wells. After incubation wells are washed with wash buffer and Anti-CRM antibody is added. After incubation, again wells are washed and Anti-Mouse IgG HRP conjugate is added. Complex is formed. After washing microwells in order to remove any non-specific binding, the substrate solution (TMB)
is added to microwells and color develops proportionally to the amount of Diptheria Toxin CRM197 in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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