Lysosomal Associated Membrane Protein 2 (LAMP2) BioAssay™ ELISA Kit (Human) is a sandwich enzyme immunoassay for in vitro quantitative measurement of LAMP2 in human serum, plasma and other biological fluids.
Detection Range
62.5-4000pg/ml
Test Principle
The microtiter plate provided in this kit has been pre-coated with an antibody specific to LAMP2. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to LAMP2. 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 LAMP2, 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 LAMP2 in the sample is then determined by comparing the O.D. of the sample to the standard curve.
Kit Components
*026602A: Microtiter Plate, 1x96 wells, Pre-coated, ready to use *026602B: Standard, 2x1 vial 026602C: Standard Diluent, 1x20ml *026602D: Detection Reagent A, 1x120ul *026602E: Detection Reagent B, 1x120ul 026602F: Assay Diluent A, 1x12ml 026602G: Assay Diluent B, 1x12ml 026602H: TMB Substrate, 1x9ml 026602J: Positive Control, 4.33ng/ml 1x1 vial 026602K: Stop Solution, 1x6ml 026602L: Wash Buffer, 30X, 1x20ml
Precaution
The Stop Solution (026602K) suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
Storage and Stability
Store *026602A, *026602B, *026602D and *026602E 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.
Assay Procedure Summary
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 TMB Substrate. Incubate 15-25 minutes at 37°C. 8. Add 50ul Stop Solution. Read at 450nm immediately.