Antidiuretic Hormone (ADH) BioAssay(TM) ELISA Kit (Human)

Product Overview

• Article Name: Antidiuretic Hormone (ADH) BioAssay(TM) ELISA Kit (Human)
• Biozol Catalog Number: USB-152290
• Supplier Catalog Number: 152290
• Alternative Catalog Number: USB-152290-48,USB-152290-96
• Manufacturer: US Biological
• Category: Kits/Assays

Product Description

The Human Antidiuretic Hormone (ADH) ELISA Kit is a competitive inhibition immunoassay for the in vitro quantitative measurement of ADH in human serum, plasma, tissue homogenates, cell lysates, cell culture supernatants and other biological fluids. Detection Range: 12.35-1000pg/ml Sensitivity: <4.71pg/ml Precision: Intra-Assay CV: <10% Inter-Assay CV: <12% 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. Test Principle: This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific for ADH has been pre-coated onto a microplate. A competitive inhibition reaction occurs between biotin-labeled ADH and unlabeled ADH (standards or samples) with the pre-coated antibody specific for ADH. 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 ADH in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of ADH in the sample. Kit Components: *152290A: Microtiter Plate, 96 wells, Pre-coated, ready to use *152290B: Standard, 2x1vial 152290C: Standard Diluent, 1x20ml *152290D: Detection Reagent A, 1x1vial *152290E: Detection Reagent B, 1x120ul 152290F: Assay Diluent A, 1x12ml 152290G: Assay Diluent B, 1x12ml 152290H: TMB Substrate, 1x9ml 152290K: Stop Solution, 1x6ml 152290L: Wash Buffer, 30X, 1x20ml 152290M: Reagent Diluent, 1x300ul Storage and Stability: Store *152290A, *152290B, *152290D and *152290E 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. Tissue Homogenates: The preparation of tissue homogenates will vary depending upon tissue type. For this assay, tissues were rinsed thoroughly in ice-cold PBS (0.02M pH 7.0-7.2) to remove excess blood and weighed before homogenization. The tissues were minced to small pieces and homogenized in 5-10ml of PBS with a glass homogenizer on ice (Micro Tissue Grinders works too). The resulting suspension was sonicated with an ultrasonic cell disrupter or subjected to two freeze-thaw cycles to further break the cell membranes. After that, the homogenates were centrifuged for 5 minutes at 5000*g. The supernatant was removed and assayed immediately or aliquoted and stored at -20C or lower. Cell Lysa