Total GLP-1 enzyme free kit instruction manual

【expected usage】

The High Sensitivity ELISA (Enzyme Linked Immunosorbent Assay) kit is designed to quantify glucagon-like peptide -1 ( 7-36 ) [GLP-1 ( 1-36 ) ] and ( 9- 36 ) The total concentration of [GLP-1(9-36)] . The basic amino acid sequence of the mammalian GLP-1 peptide chain is identical, such as: mouse, rat, pig, dog, monkey, human, and the like. The kit is for research.

[Experimental principle]

The purpose of the design, development and production of this ELISA is to quantify the active GLP-1 ( 7-36 ) and GLP-1 (9-36) in serum samples . It is based on the "two-site sandwich" technique that uses two GLP-1 specific antibodies to bind to two sites.

Standards, controls, and samples to be tested are added to streptavidin-coated microplates. Subsequently, a biotin-conjugated GLP-1 specific antibody and a HRP- labeled GLP-1 specific antibody mixture were added to each well. After the first incubation period, a "double-site sandwich" structure of "streptavidin- biotin antibody -GLP-1 ( 7-36 ) /(9-36)-HRP- labeled antibody" was formed, which The composite is adsorbed by the inner wall of the microplate. The free HRP- labeled antibody and buffer matrix are washed away during the washing process. The microplate was added with a peroxidase substrate ( 3,3',5,5' -tetramethylbenzidine, TMB ) and after a period of reaction, the absorbance was measured with a microplate reader. The ligase activity of the immunocomplex on the inner wall of the microplate and GLP-1 ( 7-36 ) / (9-36) is directly proportional to the total amount of GLP-1 in the sample .

Brief experimental steps

• Add 100 μL of standard, control and patient samples to each designated well ;
• Add 100 μL of antibody mixture to each well ;
• Static culture at 20-8 °C for 20-24 hours;
• Wash the strip with a wash buffer dilution;
• Add 200 μL of TMB matrix to each well ;
• Allow to stand at room temperature for 20 minutes;
• Add 50 μL of stop solution;
• The absorbance at 450 nm / 620 nm and 450 nm / 650 nm was read .

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