Invitrogen™ Human HIF1A ELISA Kit

Description

ELISA

The Human HIF1A ELISA quantitates Hu HIF1A in human serum, plasma, or cell culture lysates. The assay will exclusively recognize both natural and recombinant Hu HIF1A. Principle of the method The Human HIF1A solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen. Rigorous validation Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

HIF1-alpha (HIF1A) is a subunit of HIF1, which is a transcription factor found in mammalian cells cultured under reduced oxygen tension. HIF-1 is a heterodimer consisting of an alpha and beta subunit, both belonging to the basic-helix-loop-helix Per-aryl hydrocarbon receptor nuclear translocator-Sim (PAS) family of transcription factors. HIF1 functions as a transcriptional regulator of the adaptive response to hypoxia. Under hypoxic conditions, HIF-1 activates the transcription of over 40 genes, including erythropoietin, glucose transporters, glycolytic enzymes, vascular endothelial growth factor, HILPDA, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. HIF1-alpha regulates hypoxia-mediated apoptosis, cell proliferation and tumor angiogenesis. Hypoxia which induces p53 protein accumulation, directly interacts with HIF1-alpha and reduces hypoxia-induced expression of HIF1-alpha by promoting MDM2-mediated ubiquitination and proteasomal degradation under hypoxic conditions. Recent studies suggest that induction of NOX4 by HIF1-alpha contributes to maintain ROS levels after hypoxia and hypoxia-induced proliferation. In humans, it is located on the q arm of chromosome 14. The C-terminal of HIF1A binds to p300. p300/CBP-HIF complexes participate in the induction of hypoxia-responsive genes, including VEGF. Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease.

Specifications

• Accession Number: Q16665
• Assay Range: 81.92 to 20,000 pg/mL
• Assay Sensitivity: 30 pg/mL
• Description: HIF-1 Alpha ELISA Kit, 96 Assays, 1 Kit, Human Species, Immunology Research Category, Colorimetric Detection Method, Colorimetric Elisa (TMB Substrate) in Clear 96-Well Strip Plate, Contain Precoated 96-Well Strip Plate(S) and Lot-Optimized Reagents
• Assay: 96
• Conjugate: HRP
• Detection Method: Colorimetric
• Immunoassay Kit Format: Colorimetric ELISA (TMB substrate) with Plates
• Product Type: ELISA
• Protein Family: Transcription factors
• Sample Type: Cell Lysates, Plasma, Serum
• Specificity: Specific to human HIF-1 A and shows cross-reactivity with mouse and rat HIF-1 A
• Substrate Type: TMB
• Sufficient For: 96 Assays
• Target Species: Rat, Mouse
• For Use With (Application): Protein Assays and Analysis
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene ID (Entrez): 3091
• Gene Symbol: HIF-1-alpha,HIF-1 A,HIF-1 alpha,HIF1,HIF1-ALPHA,MOP1,PASD8,bHLHe78
• Host Species: Human
• Interassay CV: <10%
• Intraassay CV: <10
• Kit Contents: Pre-coated 96 well plate, Standard, Assay Diluent concentrate, Biotinylated Detection Antibody, SAV-HRP, Wash Buffer, Chromogen, Stop Solution, Adhesive Plate Covers
• Label or Dye: HRP
• Measurement: Absorbance at 450 nm minus absorbance (at 550 nm)
• Measuring Range: 450 to 550 nm
• Precision: Intra-assay CV <10%; inter-assay CV <10%
• Quantity: 96 Tests
• Regulatory Status: RUO
• Research Discipline: Immunology
• Sample Volume: Cell Lysate 50 μL, Plasma 50 μL, Serum 50 μL
• Storage Requirements: 2°C to 8°C
• Target: HIF1A
• Test Time: 1 hrs 20 mins
• Total Assay Time: 4 hrs