| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | C-P4H alpha III|collagen prolyl 4-hydroxylase alpha(III)|procollagen-proline; 2-oxoglutarate 4-dioxygenase (proline 4-hydroxylase); alpha polypeptide III|prolyl 4-hydroxylase; alpha III subunit |
| Product Type | |
| Sensitivity | |
| UniProt # |
Product Overview
Sandwich ELISA KitP4HA3 encodes a component of prolyl 4-hydroxylase, a key enzyme in collagen synthesis composed of two identical alpha subunits and two beta subunits. P4HA3 has all residues necessary to bind the peptide substrate, to coordinate Fe(2+) and the 2-oxoglutarate cosubstrate, and for enzymatic activity. Processed P4HA3 contains 525 amino acids and shares 91% and 94% amino acid identity with rat and mouse P4ha3, respectively, and 35% and 37% identity with P4HA1 and P4HA2, respectively. Northern blot an…
Assay Principle
This assay employs a two-site sandwich ELISA to quantitate P4HA3 in samples. An antibody specific for P4HA3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyP4HA3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for P4HA3 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avid…
Performance Data
| Assay Type | Sandwich ELISA |
|---|---|
| Detection Method | Sandwich ELISA |
| Detection Range | 0.156-10 ng/mL |
Stability & Storage
The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Bio…
Safety & Handling
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This is a sandwich ELISA kit. It employs a capture antibody pre-coated on the microplate surface and a biotin-conjugated detection antibody. After incubation with streptavidin-HRP and TMB substrate, a colorimetric signal proportional to analyte concentration is generated and measured at 450 nm.
The linear detection range of this kit is 0.156-10 ng/mL. Samples exceeding the upper limit must be diluted prior to testing. Samples below the lower limit may require concentration steps. Always verify that interpolated values fall within the linear portion of the standard curve.
This kit is validated for use with serum and plasma samples. Cell culture supernatants and tissue homogenates may also be tested after appropriate preparation. Avoid grossly hemolyzed, lipemic, or bacterially contaminated samples, as these may produce inaccurate results.
A microplate reader capable of measuring absorbance at 450 nm (with a 540–570 nm reference wavelength) is required. A plate washer is recommended for consistent washing steps. Calibrated micropipettes, incubator (37°C), and standard laboratory consumables are also needed.
The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O… Prepare working reagents fresh for each run. Do not freeze kit components unless specifically directed in the insert.
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