The Hycult EcHECk ELISA kit is a serological assay for the standardized detection of IgM antibodies against Enterohemorrhagic Escherichia coli (EHEC, or Shiga-toxin producing Escherichia coli; STEC) O157:H7 LPS. This is a sandwich-type ELISA kit, provided by Diapharma Group.
The kit is supplied in a standard 96-well format with removable strips. Strips are pre-coated with LPS recognizing IgM EHEC O157:H7. Biotinylated tracer antibody binds to any captured human-anti-EHEC O157:H7 IgM from the sample. Streptavidin-peroxidase conjugate binds to the biotinylated tracer antibody. Streptavidin-peroxidase conjugate then reacts with the substrate, tetramethylbenzidine (TMB). The enzyme reaction is stopped by the addition of oxalic acid. Assay absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the EcHECk standards (log). The IgM EHEC O157:H7 concentration of samples, which are run concurrently with the standards, can then be determined from the standard curve.
| Kit Component | Quantity |
| Wash Buffer, 20x | 1 vial (60 mL) |
| Dilution Buffer, 10x | 1 vial (30 mL) |
| Standard | 3 vials, lyophilized |
| Trace, biotinylated | 1 vial, 1 mL lyophilized |
| Streptavidin-peroxidase | 1 tube, 0.25 mL in solution |
| TMB substrate | 1 vial, (22 mL) |
| Stop solution | 1 vial, (22 mL) |
| 12 Microtiter strips, pre-coated | 1 plate |
Assay Time: 3.5 hours
Standard Range: 1.6 – 100 AU/mL
Detection Level: 1.6 AU/mL
Species: Human
Sample type: Serum, plasma
Storage & Stability: Product should be stored at 4ºC. Under recommended storage, product is stable for at least 6 months.
