Luminex® Discovery Assay - Human Premixed Multi-Analyte Kit
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Luminex® Discovery Assay Human Premixed Multi-Analyte Kit Catalog Number LXSAHM For the simultaneous detection of multiple human biomarkers in cell culture supernates, serum, and plasma. This package insert must be read in its entirety before using this product. For research use only. Not for use in diagnostic procedures.

TABLE OF CONTENTS SECTION Manufactured and Distributed by: USA R&D Systems, Inc. 614 McKinley Place NE, Minneapolis, MN 55413 TEL: 800 343 7475 612 379 2956 FAX: 612 656 4400 E-MAIL: info@bio-techne.com Distributed by: Europe | Middle East | Africa Bio-Techne Ltd. 19 Barton Lane, Abingdon Science Park Abingdon OX14 3NB, UK TEL: +44 (0)1235 529449 FAX: +44 (0)1235 533420 E-MAIL: info.emea@bio-techne.com China Bio-Techne China Co., Ltd. Unit 1901, Tower 3, Raffles City Changning Office, 1193 Changning Road, Shanghai PRC 200051 TEL: +86 (21) 52380373 (400) 821-3475 FAX: +86 (21) 52371001...

INTRODUCTION This kit contains the components required to screen up to 50 human biomarkers in cell culture supernate, serum, and plasma samples in multiplexed sandwich ELISAs. Luminex® Discovery Assays can be used to assess the levels of biomarkers of your choosing in a single sample. For ease of use, the microparticles are premixed in one vial as are the biotinylated detection antibodies. PRINCIPLE OF THE ASSAY Luminex® Discovery Assay multiplex kits are designed for use with any Luminex® analyzer including the MAGPIX®, Luminex® 100/200™, FLEXMAP 3D®, xMAP INTELLIFLEX®, or Bio-Rad®...

LIMITATIONS OF THE PROCEDURE • FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. • The kit should not be used beyond the expiration date on the kit label. • Do not mix or substitute reagents with those from other lots or sources. • If samples generate values higher than the highest standard, further dilute the samples with calibrator diluent and repeat the assay. • Any variation in diluent, operator, pipetting technique, washing technique, incubation time or temperature, and kit age can cause variation in binding. • Variations in sample collection, processing, and storage may...

MATERIALS PROVIDED & STORAGE CONDITIONS Store the unopened kit at 2-8 °C. Do not use past kit expiration date. This kit contains sufficient materials to run multiplex assays on one 96 well plate. PART PART # Human Standard 893899 Cocktail A+ Human Standard 893901 Cocktail B+ Human Standard 894368 Cocktail C Human Standard 893985 Cocktail D+ Human Standard 893986 Cocktail E+ Human Standard 894162 Cocktail F+ Human Standard 894625 Cocktail G+ Human Standard 896017 Cocktail H Human Standard 896018 Cocktail P Human Standard 896019 Cocktail J+ Human Standard 894824 Cocktail K Human Standard...

Premixed Microparticle Cocktail** Human Premixed 893988 Biotin-Ab Cocktail Streptavidin-PE 893535 0.6 mL of a concentrated biotinylated antibody cocktail with preservatives. 0.250 mL of a concentrated streptavidin-phycoerythrin conjugate with preservatives. 11 mL of a buffered protein base with preservatives. 2 vials (21 mL/vial) of a buffered protein base with preservatives. 21 mL of a 25-fold concentrated solution of buffered surfactant with preservative. May turn yellow over time. 1 flat-bottomed 96 well microplate used as a vessel for the assay. 1 sheet listing the selected analytes...

Note: Additional reagents supplied if CCL5/RANTES are ordered. STORAGE OF OPENED, DILUTED, OR RECONSTITUTED MATERIAL May be stored for up to 1 month at 2-8 °C.* Prepare fresh 1Xsolutions at the time of assay. Discard after use. PART PART # DESCRIPTION Human CCL5/RANTES 899028 0.075 mL a concentrated microparticle stock Magnetic Microparticle with preservatives. *Provided this is within the expiration date of the kit. SAMPLE COLLECTION & STORAGE The sample collection and storage conditions listed below are intended as general guidelines. Sample stability has not been evaluated. Cell Culture...

SAMPLE PREPARATION Use polypropylene tubes. To determine the appropriate dilution for each analyte, refer to the table located in the following link http://www.RnDSystems.com/Products/LXSAHM. Note: On the day of the assay, ALL fresh and previously frozen serum and plasma samples require centrifugation at 16,000 x g for 4 minutes immediately prior to use or dilution. Cell culture supernate, serum and plasma samples require at least a 2-fold dilution. A suggested 2-fold dilution is 75 μL of sample + 75 μL of Calibrator Diluent RD6-52. Mix thoroughly. High abundance biomarkers may require...

REAGENT PREPARATION Bring all reagents to room temperature before use. Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix gently until the crystals have completely dissolved. Add 20 mL of Wash Buffer Concentrate to 480 mL of deionized or distilled water to prepare 500 mL of Wash Buffer. Standards - Refer to the Certificate of Analysis for reconstitution volumes and assigned values of combined standard. The standards provided in the kit will differ depending on the analytes selected, but may include up to 10 unique Standard Cocktails (A-Z,1-7)....

REAGENT PREPARATION CONTINUED Pipette 200 pL of Calibrator Diluent RD6-52 into each of 5 test tubes labeled 2-6. Use Standard 1 to produce a 3-fold dilution series (below). Mix each tube thoroughly before the next transfer. Standard 1 serves as the high standard. Calibrator Diluent RD6-52 serves as the blank. DILUTED MICROPARTICLE COCKTAIL PREPARATION 1. Centrifuge the Microparticle Cocktail vial for 30 seconds at 1000 x g prior to removing the cap. 2. Gently vortex the vial to resuspend the microparticles, taking precautions not to invert the vial. 3. Dilute the Microparticle Cocktail...

1. Centrifuge the Biotin-Antibody Cocktail vial for 30 seconds at 1000 x g prior to removing the cap. 2. Gently vortex the vial, taking precautions not to invert the vial. 3. Dilute the Biotin-Antibody Cocktail in Diluent RD2-1. Mix gently.