EGLN1 ELISA Kit (Human)

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%

C1orf12; ECYT3; egl nine homolog 1; egl nine-like protein 1; HALAH; HIFPH2; HIF-PH2; HIF-prolyl hydroxylase 2; HPH2; HPH-2; hypoxia-inducible factor prolyl hydroxylase 2; PHD2; prolyl hydroxylase domain-containing protein 2; SM20; SM-20; zinc finger MYND domain-containing protein 6; ZMYND6.

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.

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.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) _x005F Serum (n=5) 91-101 96 _x005F EDTA Plasma (n=5) 97-105 102 _x005F Heparin Plasma (n=5) 84-101 91 _x005F Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 91-101 96 Serum (n=5) 91-101 96 EDTA Plasma (n=5) 97-105 102 EDTA Plasma (n=5) 97-105 102 Heparin Plasma (n=5) 84-101 91 Heparin Plasma (n=5) 84-101 91

Component Amount _x005F Anti-EGLN1 Microplate 96 Wells (12 x 8 Well Strips) _x005F EGLN1 Lyophilized Standard 2 x 2 ng _x005F 100X Biotinylated EGLN1 Detector Antibody 120 uL _x005F Avidin/HRP Conjugate 120 uL _x005F Standard Diluent 1 x 20 mL _x005F Detector Antibody Diluent 1 x 12 mL _x005F Conjugate Diluent 1 x 12 mL _x005F 30X Wash Buffer 1 x 20 mL _x005F TMB Substrate 1 x 9 mL _x005F Stop Solution 1 x 6 mL _x005F Component Amount Component Amount Anti-EGLN1 Microplate 96 Wells (12 x 8 Well Strips) Anti-EGLN1 Microplate 96 Wells (12 x 8 Well Strips) EGLN1 Lyophilized Standard 2 x 2 ng EGLN1 Lyophilized Standard 2 x 2 ng 100X Biotinylated EGLN1 Detector Antibody 120 uL 100X Biotinylated EGLN1 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Egl Nine Homolog 1 (EGLN1) ._x005F No significant cross-reactivity or interference between Egl Nine Homolog 1 (EGLN1) and analogues was observed.