EPAS1 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Hypoxia Inducible Factor 2 Alpha (HIF2a) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Hypoxia Inducible Factor 2 Alpha (HIF2a) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Basic-helix-loop-helix-PAS protein MOP2; bHLHe73; class E basic helix-loop-helix protein 73; ECYT4; endothelial PAS domain-containing protein 1; EPAS-1; HIF-1alpha-like factor; HIF-1-alpha-like factor; HIF2A; HIF2-alpha; HIF-2-alpha; HLF; hypoxia-inducible factor 2 alpha; Hypoxia-inducible factor 2-alpha; member of PAS protein 2; MOP2; PAS domain-containing protein 2; PASD2.

Transcription factor involved in the induction of oxygen regulated genes. Heterodimerizes with ARNT; heterodimer binds to core DNA sequence 5'-TACGTG-3' within the hypoxia response element (HRE) of target gene promoters (By similarity) . Regulates the vascular endothelial growth factor (VEGF) expression and seems to be implicated in the development of blood vessels and the tubular system of lung. May also play a role in the formation of the endothelium that gives rise to the blood brain barrier. Potent activator of the Tie-2 tyrosine kinase expression. Activation requires recruitment of transcriptional coactivators such as CREBBP and probably EP300. Interaction with redox regulatory protein APEX seems to activate CTAD (By similarity) .

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 Hypoxia Inducible Factor 2 Alpha (HIF2a) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Hypoxia Inducible Factor 2 Alpha (HIF2a) . 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 Hypoxia Inducible Factor 2 Alpha (HIF2a), 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 Hypoxia Inducible Factor 2 Alpha (HIF2a) 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

Component Amount EPAS1 Microplate 96 Wells (12 x 8 Well strips) EPAS1 Lyophilized Standard 2 100X EPAS1 HRP-Detector Anitbody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 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 2 mL Stop Solution 1 x 20 mL Component Amount Component Amount EPAS1 Microplate 96 Wells (12 x 8 Well strips) EPAS1 Microplate 96 Wells (12 x 8 Well strips) EPAS1 Lyophilized Standard 2 EPAS1 Lyophilized Standard 2 100X EPAS1 HRP-Detector Anitbody 1 x 120 uL 100X EPAS1 HRP-Detector Anitbody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 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 2 mL TMB Substrate 1 x 2 mL Stop Solution 1 x 20 mL Stop Solution 1 x 20 mL