Enhanced chemiluminescence technique
Enhanced chemiluminescence (ECL) is a technique used for the detection and quantification of proteins in Western blot analysis. It involves the use of a chemiluminescent substrate that emits light when it reacts with the enzyme-labeled target protein. This technique provides a sensitive and reliable method for protein visualization and analysis.
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7 protocols using enhanced chemiluminescence technique
Hypoxia Regulation of Proliferation and Angiogenesis
Immunoprecipitation and Western Blot Analysis
Uncropped scans of all blots are provided as
Endothelial Cell Surface Biotinylation
Co-immunoprecipitation and Western Blotting of STIM1 and Dynein
Cellular lysates were incubated for 20 min on wet ice and then centrifuged at 15,000 g, 20 min, at 4°C. The total protein amount was determined using the bicinchoninic acid (BCA) assay (Pierce). Equivalent amounts (1 mg) of protein were immunoprecipitated for 2 h at 4°C with the antibody of interest, and immune complexes were recovered on protein A‐ or G‐Sepharose (GE Healthcare) for 1 h at 4°C. Where indicated in the figure legend, cell lysates were pre‐cleared on Sepharose beads before incubation with the antibody of interest. Controls of immunoprecipitations were performed incubating cell lysate with empty immunoglobulin (Ig) G of the same specie of the antibody of interest (Normal rabbit, NI#01, or mouse, NI#03, IgG from Merck Life Science). Immunoprecipitates were washed four times with lysis buffer with or without detergent and then separated by SDS–PAGE. Proteins were then transferred to a Hybond‐C extra nitrocellulose membrane (Amersham), probed with antibodies of interest, and detected by enhanced chemiluminescence technique (PerkinElmer).
Endogenous LPHN2 Immunoprecipitation and Western Blot
Protein Extraction and Analysis via SDS-PAGE
HIF-1α Protein Expression Analysis
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