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广域小鼠胰岛素酶免试剂盒

Cat NoBIOK1998
Conjugate
Type胰岛素含量测定
SourceN/A
Size96T
ApplicationELISA
FormatLiquid
SpeciesMouse
StorageStore at 2-8°C upon receipt. Remove any unused antibody\u0002coated strips from the microplate, return them to the foil pouch and re-seal.
Once opened, the strips may be stored at 2-8°C for up to one month.
SynonymsMouse insulin elisa kit
Purification
NoteAll reagents and micro-plate strips should be equilibrated to room
temperature prior to use. A standard curve must be performed with each
assay. It is recommended that all standards and samples should be run in
duplicate.Research Use Only. Not for Use in Diagnostic Procedures.
MolecularWeight
DescriptionPreparation of reagentsA. 1×Wash buffer.Prepare 1×Wash buffer by mixing the 10×Wash buffer (20 ml) with 180 ml of distilled water or deionized water. The 1×Wash buffer may be stored at 2-8°C for up to one month.B. 1×Detection antibody solution.Prepare 1×Detection antibody solution by dilution of the 100×Detection antibody solution in Assay buffer, mix well. 70 μl of the 1×Detection antibody solution is required per well. Preparation of samplesIf a sample has a greater concentration of insulin than the highest standard, the sample should be diluted with 0 ng/ml insulin standard solution and the assay should be repeated.Assay procedure1. Add 70 μl of 1x Detection antibody solution to each well.2. Add 5 μl of standard or sample to its respective well.3. Cover the plate with a plate sealer. Incubate at room temperature for 1 hour, shaking the plate at 600 rpm on a horizontal micro-plate shaker. (*Alternative incubation step in the absence of shaker: gently tap the plate frame for a few seconds to ensure thorough mixing, incubate at room temperature for 1.5 hours.\u00024. Discard well contents and remove any remaining solution by inverting and tapping the plate on a clean paper towel. Add 300 μl of 1× Wash buffer to each well. Incubate at room temperature for 30 seconds.5. Discard the 1× Wash buffer and tap the plate on a clean paper towel to remove residual wash buffer. Repeat the wash step for a total 4 washes.6. Add 100 μl of Substrate solution to each well, incubate at room temperature for 15 minutes. Protect from light.7. Add 100 μl of Stop solution to each well, gently tap the plate frame for a few seconds to ensure thorough mixing. Measure absorbance of each well at 450 nm immediately. Calculation1. Subtract the absorbance of the blank from that of standards and samples.2. Generate a standard curve by plotting the absorbance obtained (y-axis) against insulin concentrations (x-axis) . The best fit line can be generated with any curve-fitting software by regression analysis. Log-log curve\u0002fitting is recommended for data analysis. 3. Determine insulin concentration of samples from standard curve.
BackgroundThis assay is a sandwich ELISA based on two monoclonal antibodies against separate antigenic determinants on the insulin molecule.
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