Coagulation Factor II (F2) BioAssay(TM) ELISA Kit (Mouse)

The Mouse Coagulation Factor II (F2) ELISA kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of F2 in mouse plasma. Detection Range: 1.56-100ng/ml, Sensitivity: <0.59ng/ml, Precision: Intra-Assay: CV<10%, Inter-Assay: CV<12%, Kit Components: *024221A: Microtiter Plate, 1x96 wells, Pre-coated, ready to use., *024221B: Standard, 2x1vial , 024221C: Standard Diluent, 1x20ml, *024221D: Detection Reagent A, 1x120ul , *024221E: Detection Reagent B, 1x120ul , 024221F: Assay Diluent A, 1x12ml, 024221G: Assay Diluent B, 1x12ml, 024221H: TMB Substrate, 1x9ml, 024221K: Stop Solution, 1x6ml, 024221L: Wash Buffer, 30X, 1x20ml , Precaution: The Stop Solution (024221K) 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 *024221A, *024221B, *024221D and *024221E 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. Materials Required But Not Supplied: 1. Microplate reader with 450 ± 10nm filter., 2. Precision single or multi-channel pipettes and disposable tips., 3. Eppendorf Tubes for diluting samples., 4. Deionized or distilled water., 5. Absorbent paper for blotting the microtiter plate., 6. Container for Wash Solution, Sample Preparation and Storage: Plasma:, Collect plasma using 3.8% sodium citrate (sodium citrate : blood = 1 : 9) as an anticoagulant. Centrifuge samples for 15 minutes at 1000xg at 2-8°C within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquots at -20°C or -80°C for later use. Avoid repeated freeze/thaw cycles. Note: Plasma samples require about 2,000-fold dilution. A serial dilution of the sample can be performed. Prepare a 1:20 dilution by transferring 20ul sample to 380ul PBS. Next, dilute further by transferring 10ul of the 1:20 diluted sample to 990ul PBS. This yields a 1:2,000 dilution of the sample. Mix contents thoroughly at each stage of the dilution process. Samples should be diluted using 0.02M PBS, pH 7.0-7.2. , 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.