The Measles Virus (MV) Antibody (IgG) BioAssay(TM) ELISA Kit (Human) is for the qualitative determination of MV antibody (IgG) concentration in human serum. This kit is intended for research use only and may not be used for therapeutic or diagnostic applications Principle of the Test: This assay employs the qualitative enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with antigen. Samples are pipetted into the wells with anti-human IgG conjugated Horseradish Peroxidase (HRP). Any antibodies specific for the antigen present will bind to the pre-coated antigen. Following a wash to remove any unbound reagent, a substrate solution is added to the wells and color develops in proportion to the amount of human MV antibody (IgG) bound in the initial step. The color development is stopped and the intensity of the color is measured. Kit Components: 351664A: Microtiter Strips, 1x96 wells. Pre-coated. 351664B: Negative Control, 1x300ul 351664C: Positive Control, 1x300ul 351664D: Sample Diluent, 1x12ml 351664E: HRP Conjugate, 1x12ml 351664F: Wash Buffer (30X), 1x30ml 351664G: Substrate A, 1x6ml 351664H: Substrate B, 1x6ml 351664J: Stop Solution, 1x6ml Storage and Stability: Store kit at 4C. The unopened kit is stable for 6 months after receipt. Once opened, the kit is stable for up to 2 weeks at 4C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Sample Collection and Storage: Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at RT or overnight at 4C before centrifugation for 15 minutes at 1000xg. Remove serum and assay immediately or aliquot and store samples at -20C or -80C. Avoid repeated freeze-thaw cycles. Serum samples require a 40-fold dilution into normal saline before test. The suggested 40-fold dilution can be achieved by adding 10ul sample to 390ul of normal saline. Assay Procedure: Bring all reagents and samples to RT before use. Centrifuge the sample again after thawing before the assay. It is recommended that all samples and controls be assayed in duplicate. 1. Prepare all reagents, and *samples as directed in the previous sections. 2. Determine the number of wells to be used and put any remaining wells and the desiccant back into the pouch and seal the ziploc, store unused wells at 4C. 3. Set a Blank well with 100ul of Sample Diluent. 4. Add 100ul Sample Diluent to per well. Then add 10ul of Negative Control, Positive Control or Sample per well. 5. Cover with the adhesive strip provided. Incubate for 45 minutes at 37C. 6. Aspirate each well and wash, repeating the process three times for a total of four washes. Wash by filling each well with 200ul of 1X Wash Buffer using a squirt bottle, multi-channel pipette, manifold dispenser or autowasher, and let it stand for 30 seconds, complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining 1X Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels. 7. Add 100ul of HRP Conjugate to each well (not to Blank!). Cover the microtiter plate with the adhesive strip. Incubate for 30 minutes at 37C. 8. Repeat the aspiration/wash process for four times as in step 6. 9. Add 50ul of Substrate A and 50ul Substrate B to each well. Incubate for 10 minutes at 37C. Protect from light. 10. Add 50ul of Stop Solution to each well, gently tap the plate to ensure thorough mixing. 11. Determine the optical density of each well within 10 minutes using a microplate reader set to 450nm. Take blank well as zero. *Samples may require dilution. Contact: United States Biological 4 Technology Way Salem, MA 01970 www.usbio.net Tech Support: tech usbio.net Customer Service: service usbio.net Tel: 781-639-5092 800-520-3011 Fax: 978-594-8052
* VAT and and shipping costs not included. Errors and price changes excepted