1.23-100ug/ml

0.53ug/ml

This assay has high sensitivity and excellent specificity for detection of 2,3-Bisphosphoglycerate (2,3-BPG). No significant cross-reactivity or interference between 2,3-Bisphosphoglycerate (2,3-BPG) and analogues was observed.

Intra-Assay %CV: <10% Inter-Assay %CV: <12%

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to 2,3-BPG has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled 2,3-BPG and unlabeled 2,3-BPG (Standards or samples) with the pre-coated antibody specific to 2,3-BPG. 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 2,3-BPG in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of 2,3-BPG in the sample.

Pre-coated, ready to use 96-well strip plate Standard, 2x vials Detection Reagent A, 1x120ul Detection Reagent B, 1x120ul TMB Substrate, 1x9ml Wash Buffer (30X): 1x20ml Standard Diluent, 1x20ml Assay Diluent A, 1x12ml Assay Diluent B, 1x12ml Stop Solution, 1x6ml

Store Microtiter Strips, Standard, Detection Reagents and Positive Control at -20°C. After reconstitution of Positive 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.

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 Substrate Solution. Incubate 10-20 minutes at 37°C. 8. Add 50ul Stop Solution. Read at 450nm immediately.