Hypoxia-inducible factor-1α (HIF-1α) Test Kit – User Instruction
Objective: This kit is designed for the quantitative determination of hypoxia-inducible factor-1α (HIF-1α) in rat serum, plasma, and other biological fluid samples. It provides a reliable and efficient method to assess HIF-1α levels, which are closely associated with cellular responses to hypoxia and various pathological conditions.
Principle of the Assay: The HIF-1α test is based on the double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) technique. A microtiter plate is pre-coated with a specific monoclonal antibody against rat HIF-1α. After adding the sample, HIF-1α present in the sample binds to the immobilized antibody. Subsequently, a horseradish peroxidase (HRP)-labeled secondary antibody is introduced, forming a complex of antibody-HIF-1α-enzyme-labeled antibody. Following a washing step to remove unbound components, a TMB substrate solution is added. The enzyme catalyzes the conversion of TMB into a blue product, which changes to yellow in the presence of an acidic stop solution. The intensity of the color is directly proportional to the concentration of HIF-1α in the sample. The absorbance is measured at 450 nm using a microplate reader, and the HIF-1α concentration is determined by comparing the OD values to a standard curve generated from known concentrations.
This method ensures high specificity, sensitivity, and reproducibility, making it suitable for research applications involving hypoxia-related studies and biomarker analysis.
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