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Human Transmembrane prolyl 4-hydroxylase (P4HTM) ELISA Kit

Due to the possibility of mismatching between antigens from other origin and antibodies used in our kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.

Principle of the Assay

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 Human P4HTM. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human P4HTM. 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 Human P4HTM, 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 Human P4HTM in the samples is then determined by comparing the OD of the samples to the standard curve.

For use with: Serum, plasma, tissue homogenates, cell lysates, cell culture supernates, and other biological fluids.

For Research Use Only. Not for use in diagnostic procedures.

Target Information

SWISS PROT Q9NXG6
SYNONYMS EGLN4; FLJ20262; HIF prolyl hydroxylase PH4; HIF-PH4; HIF-prolyl hydroxylase 4; HIFPH4; HPH-4; Hypoxia inducible factor prolyl 4 hydroxylase; Hypoxia inducible factor prolyl hydroxylase; Hypoxia-inducible factor prolyl hydroxylase 4; P4H TM; P4H with transmembrane domain; P4H-TM; P4htm; P4HTM_HUMAN; PH 4; PH4; PHD4; Proline 4 hydroxylase; Prolyl 4 hydroxylase transmembrane (endoplasmic reticulum); Prolyl hydroxlase domain containing 4; Transmembrane prolyl 4-hydroxylase

Materials Supplied

Kit Components 96 Wells Quantity/Size
Pre-Coated Microplate 48T: 6 strips x 8 wells; 96T: 12 strips x 8 wells
Standard (Lyophilized) 48T: 1 vial; 96T: 2 vials
Biotinylated Antibody (100×) 48T: 60 μL; 96T: 120 μL
Streptavidin-HRP (100×) 48T: 60 μL; 96T: 120 μL
Standard/Sample Diluent Buffer 48T: 10 mL; 96T: 20 mL
Biotinylated Antibody Diluent 48T: 6 mL; 96T: 12 mL
HRP Diluent 48T: 6 mL; 96T: 12 mL
Wash Buffer (25×) 48T: 10 mL; 96T: 20 mL
TMB Substrate Solution 48T: 6 mL; 96T: 10 mL
Stop Reagent 48T: 3 mL; 96T: 6 mL
Plate Covers 48T: 1 piece; 96T: 2 pieces

Storage

Storage *If the kit is opened, Store the whole kit at 4°C. If the kit is not used up in 1 week. Store the Pre-Coated Microplate, Standard ,Biotinylated Antibody and Streptavidin-HRP at -20°C, the rest reagents at 4°C, please used up within 6 months. *If the kit is not opened, store the whole kit: 4°C(short time storage, valid for 6 months); -20°C (long-term storage, valid for 1 year). Avoid repeated freeze-thaw cycles.

Performance Characteristics

REPEATABILITY Intra-assay Precision (Precision within an assay):CV% < 8% Three samples of known concentration were tested twenty times on 1 plate to assess intra-assay precision. Inter-assay Precision (precision between assays):CV% < 10% Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
SENSITIVITY The minimum detectable dose was 5.5pg/mL.
ASSAY RANGE 15.63–1,000pg/mL
SPECIFICITY This assay has high sensitivity and excellent specificity for detection of Human P4HTM. No significant cross-reactivity or interference between Human P4HTM and analogues was observed.