Tryptophan (Trp) BioAssay™ ELISA Kit (General)

The Tryptophan (Trp) BioAssay™ ELISA Kit utilizes the competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of tryptophan in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.||Detection Range:|1.23-100ug/ml||Sensitivity:|0.55ug/ml||Intra-Assay: |CV<10% ||Inter-Assay: |CV<12%||Assay Principle:|A monoclonal antibody specific to tryptophan has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled tryptophan and unlabeled tryptophan (Standards or samples) with the pre-coated antibody specific to tryptophan. 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 reverse proportional to the concentration of tryptophan in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of tryptophan in the sample.||Kit Components:|Microtiter Strips, 1x96wells, Pre-coated, ready to use|Standard, 2x1vial |Standard Diluent, 1x20ml|Detection Reagent A, 1x120ul |Detection Reagent B, 1x120ul |Assay Diluent A, 1x12ml|Assay Diluent B, 1x12ml|TMB Substrate, 1x9ml|Positive Control, 1x1vial (Lyophilized)|Stop Solution, 1x6ml|Wash Buffer, 30X, 1x20ml||Storage and Stability:|Store microtiter strips, Standard, Detection Reagent A, Detection Reagent B and positive control at -20°C. After reconstitution of the postive control store at 4°C and use within 5 days. 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 100ul standard or sample to each well. Incubate 1 hour 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 10-20 minutes at 37°C.|8. Add 50ul Stop Solution. Read at 450nm immediately.