The Diapharma Human C3a ELISA kit 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 antibody to capture C3a in the sample. Biotinylated tracer antibody binds to captured C3a. 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. The 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 C3a standards (log). The C3a concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.

Assay Overview:

 

Wash/Dilution Buffer, 20x — 1 vial (60 ml)

Sample Dilution Buffer, 10x — 1 vial (20 ml)

Standard — 2 vials, lyophilized

Tracer, biotinylated — 1 vial, 1 ml lyophilized

Streptavidin-peroxidase, 100x — 1 tube, 0.25 ml in solution

TBM Substrate — 1 vial (11 ml)

Stop Solution  — 1 vial (22 ml)

12 pre-coated Microtiter well strips — 1 plate

Assay Time: 3.5 hours

Standard Range: 31.3 – 2000 pg/ml

Detection Level: 31.3 pg/ml

Standard: Native human protein

Species: Human

Sample type: Serum, Plasma, BAL fluid, urine