EGLN1 ELISA Kit (Human)

ELISA Kit

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EGLN1

426

Cellular oxygen sensor that catalyzes, under normoxic conditions, the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates a specific proline found in each of the oxygen-dependent degradation (ODD) domains (N-terminal, NODD, and C-terminal, CODD) of HIF1A. Also hydroxylates HIF2A. Has a preference for the CODD site for both HIF1A and HIF1B. Hydroxylated HIFs are then targeted for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. Under hypoxic conditions, the hydroxylation reaction is attenuated allowing HIFs to escape degradation resulting in their translocation to the nucleus, heterodimerization with HIF1B, and increased expression of hypoxy-inducible genes. EGLN1 is the most important isozyme under normoxia and, through regulating the stability of HIF1, involved in various hypoxia-influenced processes such as angiogenesis in retinal and cardiac functionality. Target proteins are preferentially recognized via a LXXLAP motif.

NP_071334.1

This assay has high sensitivity and excellent specificity for detection of Egl Nine Homolog 1 (EGLN1). No significant cross-reactivity or interference between Egl Nine Homolog 1 (EGLN1) and analogues was observed.

Component: Amount
Anti-EGLN1 Microplate: 96 Wells (12 x 8 Well Strips)
EGLN1 Lyophilized Standard: 2 x 2 ng
100X Biotinylated EGLN1 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 Egl Nine Homolog 1 (EGLN1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Egl Nine Homolog 1 (EGLN1). 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 Egl Nine Homolog 1 (EGLN1), 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 Egl Nine Homolog 1 (EGLN1) 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 Egl Nine Homolog 1 (EGLN1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Egl Nine Homolog 1 (EGLN1) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%