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Human P4HTM(Transmembrane prolyl 4-hydroxylase) ELISA Kit
ELK9846
Size： Price：
48 T $412.00
96 T $588.00
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Overview

Product name: Human P4HTM(Transmembrane prolyl 4-hydroxylase) ELISA Kit
Reactivity: Human
Alternative Names: 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;
Assay Type: Sandwich
Sensitivity: 5.5 pg/mL
Standard: 1000 pg/mL
Detection Range: 15.63-1000 pg/mL
Sample Type: Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Cardiovascular; Epigenetics and Nuclear Signaling; Cancer;
Uniprot ID: Q9NXG6
Test principle: 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.

Standard curve

Concentration (pg/mL) OD Corrected OD
1000.00 2.131 2.042
500.00 1.663 1.574
250.00 1.193 1.104
125.00 0.828 0.739
62.50 0.527 0.438
31.25 0.357 0.268
15.63 0.241 0.152
0.00 0.089 0.000

Precision

Intra-assay Precision (Precision within an assay)：CV%<8%

Three samples of known concentration were tested twenty times on one 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.

Recovery

Matrices listed below were spiked with certain level of recombinant P4HTM and the recovery rates were calculated by comparing the measured value to the expected amount of P4HTM in samples.
Matrix Recovery range Average
serum(n=5) 93-107% 100%
EDTA plasma(n=5) 78-90% 84%
Heparin plasma(n=5) 82-95% 88%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of P4HTM and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 81-96% 87-96% 85-99% 79-93%
EDTA plasma(n=5) 89-97% 91-102% 95-106% 85-101%
Heparin plasma(n=5) 87-102% 84-98% 83-96% 93-102%