The Human Insulin (INS) ELISA Kit is a competitive inhibition immunoassay for the in vitro quantitative measurement of INS in human serum, plasma, cell culture supernatant and other biological fluids.
Test Principle:This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific for INS has been pre-coated onto a microplate. A competitive inhibition reaction occurs between biotin-labeled INS and unlabeled INS (standards or samples) with the pre-coated antibody specific for INS. After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is inversely proportional to the concentration of INS in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of INS in the sample.
Kit Components:*025826A: Microtiter Strip, 1x96 wells, Pre-coated, ready to use*025826B: Standard, 2x1vial 025826C: Standard Diluent, 1x20ml*025826D: Detection Reagent A, 1x120ul *025826E: Detection Reagent B, 1x120ul 025826F: Assay Diluent A, 1x12ml025826G: Assay Diluent B, 1x12ml025826H: TMB Substrate, 1x9ml*025826J: Positive Control 3191.5pg/ml025826K: Stop Solution, 1x6ml025826L: Wash Buffer, 30X, 1x20ml
Storage and Stability:Store *025826A, *025826B, *025826D, *025826E and *025826J at -20°C. After reconstitution of *024351J, store at 4°C and use within 5 days. Store all the other components at 4°C. Unused kit is stable for 6 months. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.
Notes on 025826J: Positive Control:Intended Use:The Positive Control is from a human serum sample known to contain INS. It will provide a positive result in the ELISA assay, which indicates that the kit is optimized and working properly.
Characteristics:The positive control vial contains human INS in human serum (lyophilized powder). Note: The human based material used in this product was screened for HIV, Hepatitis A, B, C, HTLV and Syphilis.
Expected Values:3191.5pg/ml
Usage:Dissolve with 150ul standard dilutent and let stand for 10 minutes. If powder is partly insoluble, shake gently until fully dissolved. Mix gently before use. In the experiment, add 50ul to an appropriate well of the plate and test its concentration along with standards and your specimens. Note: Upon receipt store Positive Control vial at -20°C. After reconstitution, store Positive Control vial at 4°C and use within 5 days.
Assay Procedure Summary:1. Prepare all reagents, samples and standards
2. Add 50ul standard or sample to each well, then add 50ul prepared Detection Reagent A immediately. Shake and mix. Incubate 1 hour at 37°C
3. Aspirate and wash 3 times
4. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37°C
5. Aspirate and wash 5 times
6. Add 90ul Substrate Solution. Incubate 15-25 minutes at 37°C
7. Add 50ul Stop Solution. Read absorbance at 450nm immediately.
