12.5-800ng/ml

≤4.6ng/ml

The microtiter plate provided in this kit has been pre-coated with an antibody specific to rat IGFBP3. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for rat IGFBP3. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain IGFBP3, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of IGFBP3 in the sample is then determined by comparing the O.D. of the sample to the standard curve.

*025875A: Microtiter Plate, 1x96 wells, Pre-coated; ready to use *025875B: Standard, 2x1vial 025875C: Standard Diluent, 1x20ml *025875D: Detection Reagent A, 1x120ul *025875E: Detection Reagent B, 1x120ul 025875F: Assay Diluent A, 1x12ml 025875G: Assay Diluent B, 1x12ml 025875H: TMB Substrate, 1x9ml *025875J: Positive Control, 231.8ng/ml 1x1vial 025875K: Stop Solution, 1x6ml 025875L: Wash Buffer, 30X, 1x20ml

The Stop Solution (025875K) suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

The control vial is from a rat serum sample known to contain the target protein. It will provide a positive result in the ELISA assay, which indicates that the kit is optimized and working properly.

The control vial contains rat IGFBP3 in rat serum (lyophilized powder). Note: The rat based material used in this product is from healthy rats of SPF standard (specific pathogen-free animals).

231.8ng/ml

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 100ul to an appropriate well of the plate and test its concentration along with standards and your specimens. Note: After reconstitution, store Positive Control vial at 4°C and use within 5 days.

Store *025875A, *025875B, *025875D, *025875E and *025875J at -20°C. 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.

1. Prepare all reagents, samples and standards. 2. Add 100ul standard or sample to each well. Incubate 2 hours at 37°C. 3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37°C. 4. Aspirate and wash 3 times. 5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37°C. 6. Aspirate and wash 5 times. 7. Add 90ul Substrate Solution. Incubate 15-25 minutes at 37°C. 8. Add 50ul Stop Solution. Read at 450nm immediately.