Estrone (E1) BioAssay(TM) ELISA Kit

Produktübersicht

• Artikelname: Estrone (E1) BioAssay(TM) ELISA Kit
• Artikelnummer: USB-024896
• Hersteller Artikelnummer: 024896
• Alternativnummer: USB-024896-96
• Hersteller: US Biological
• Kategorie: Kits/Assays

Produktbeschreibung

The Estrone (E1) ELISA Kit is a competitive inhibition immunoassay for the in vitro quantitative measurement of E1 in human serum, plasma and other biological fluids. Detection Range: 24.69-2,000pg/ml Sensitivity: <8.86pg/ml Precision: Intra-Assay CV: <10% Inter-Assay CV: <12% Test Principle: This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific for E1 has been pre-coated onto a microplate. A competitive inhibition reaction occurs between biotin-labeled E1 and unlabeled E1 (standards or samples) with the pre-coated antibody specific for E1. 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 E1 in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of E1 in the sample. Kit Components: *024896A: Microtiter Plate, 96 wells, Pre-coated, ready to use *024896B: Standard, 2x1vial 024896C: Standard Diluent, 1x20ml *024896D: Detection Reagent A, 1x1vial *024896E: Detection Reagent B, 1x120ul 024896F: Assay Diluent A, 1x12ml 024896G: Assay Diluent B, 1x12ml 024896H: TMB Substrate, 1x9ml 024896K: Stop Solution, 1x6ml 024896L: Wash Buffer, 30X, 1x20ml 024896M: Reagent Diluent, 1x300ul Storage and Stability: Store *024896A, *024896B, *024896D and *024896E at -20C. Store all the other components at 4C. 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: Serum: Allow samples to clot for two hours at room temperature or overnight at 4C before centrifugation for 20 minutes at approximately 1000xg. Assay freshly prepared serum immediately or store samples in aliquots at -20C or -80C for later use. Avoid repeated freeze/thaw cycles. Plasma: Collect plasma using EDTA or heparin as anticoagulant. Centrifuge samples for 15 minutes at 1000xg at 2-8C within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquots at -20C or -80C for later use. Avoid repeated freeze/thaw cycles. Other Biological Fluids: Centrifuge samples for 20 minutes at 1000*g. Remove particulates and assay immediately or store samples in aliquots at -20C or -80C. Avoid repeated freeze/thaw cycles. Note: 1. Samples to be used within 5 days may be stored at 4C, otherwise samples must be stored at -20C (1 month) or -80C (2 months) to avoid loss of bioactivity and contamination. 2. Sample hemolysis will influence the results so hemolyzed specimens should not be used. 3. When performing the assay, bring samples to room temperature. 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 37C. 3. Aspirate and wash 3 times. 4. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37C. 5. Aspirate and wash 5 times. 6. Add 90ul Substrate Solution. Incubate 15-25 minutes at 37C. 7. Add 50ul Stop Solution. Read absorbance at 450nm immediately.