Kallikrein 1 (KLK1) BioAssayTM ELISA Kit (Rat) is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of KLK1 in rat serum, plasma, tissue homogenates and other biological fluids.
Detection Range:123.5-10,000pg/ml
Sensitivity:48.2pg/ml
Intra-Assay CV:<10%
Inter-Assay CV:<12%
Test Principle:This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific for KLK1 has been pre-coated onto a microplate. A competitive inhibition reaction occurs between biotin-labeled KLK1 and unlabeled KLK1 (standards or samples) for limited binding sites on the pre-coated antibody specific for KLK1. 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 KLK1 in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of KLK1 in the sample.
Precaution:The Stop Solution (026320K) suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothingprotection when using this material.
Storage and Stability:Store *026320A, *026320B, *026320D and *026320E at -20°C. Store all the other components at 4°C. Unused kit is stable for 6 months after receipt. 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.
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.
