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.