EPAS1 ELISA Kit (Mouse)

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%

BHLHe7; bHLHe73; endothelial PAS domain-containing protein 1; EPAS-1; HIF; Hif like protein; HIF1 alpha-like factor; HIF1alpha-like factor; HIF-1-alpha-like factor; HIF-2; HIF2A; HIF2-alpha; HIF-2alpha; HIF-2-alpha; HIF-related factor; HL; HLF; HLF (HIF1alpha-like factor) ; HR; HRF; hypoxia inducible transcription factor 2alpha; hypoxia-inducible factor 2-alpha; mHLF; MO; MOP2.

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 (PubMed:26245371) . 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

Matrix Recovery range (%) Average (%) Serum (n=5) 85-103 96 EDTA Plasma (n=5) 80-98 94 Heparin Plasma (n=5) 79-90 83 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 85-103 96 Serum (n=5) 85-103 96 EDTA Plasma (n=5) 80-98 94 EDTA Plasma (n=5) 80-98 94 Heparin Plasma (n=5) 79-90 83 Heparin Plasma (n=5) 79-90 83

Component Amount Anti-EPAS1 Microplate 96 Wells (12 x 8 Well strips) EPAS1 Lyophilized Standard 2 100X Biotinylated EPAS1 Detector Antibody 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 Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-EPAS1 Microplate 96 Wells (12 x 8 Well strips) Anti-EPAS1 Microplate 96 Wells (12 x 8 Well strips) EPAS1 Lyophilized Standard 2 EPAS1 Lyophilized Standard 2 100X Biotinylated EPAS1 Detector Antibody 1 x 120 uL 100X Biotinylated EPAS1 Detector Antibody 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 Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

This assay has high sensitivity and excellent specificity for detection of Hypoxia Inducible Factor 2 Alpha (HIF2a) . No significant cross-reactivity or interference between Hypoxia Inducible Factor 2 Alpha (HIF2a) and analogues was observed.