HP High Sensitivity ELISA Kit (Human)

Enzyme-linked Immunosorbent assay-Sandwich

Wet Ice

haptoglobin

As a result of hemolysis, hemoglobin is found to accumulate in the kidney and is secreted in the urine. Haptoglobin captures, and combines with free plasma hemoglobin to allow hepatic recycling of heme iron and to prevent kidney damage. Haptoglobin also acts as an Antimicrobial; Antioxidant, has antibacterial activity and plays a role in modulating many aspects of the acute phase response. Hemoglobin/haptoglobin complexes are rapidely cleared by the macrophage CD163 scavenger receptor expressed on the surface of liver Kupfer cells through an endocytic lysosomal degradation pathway.

NP_001119574.1

This assay has high sensitivity and excellent specificity for detection of Haptoglobin (Hpt). No significant cross-reactivity or interference between Haptoglobin (Hpt) and analogues was observed.

Component: Amount
Anti-HP Microplate: 96 Wells (12 x 8 Well Strips)
HP Lyophilized Standard: 2
100X Biotinylated HP Detector Antibody: 120 uL
Avidin/HRP Conjugate: 120 uL
Standard Diluent: 1 x 20 mL
Detector Antibody Diluent: 1 x 12 mL
Conjugate Diluent: 1 x 12 mL
30X Wash Buffer: 1 x 20 mL
TMB Substrate: 1 x 9 mL
Stop Solution: 1 x 6 mL

3h

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to High Sensitive Haptoglobin (Hpt). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to High Sensitive Haptoglobin (Hpt). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain High Sensitive Haptoglobin (Hpt), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of High Sensitive Haptoglobin (Hpt) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Haptoglobin (Hpt) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Haptoglobin (Hpt) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%