RayPlex® Human Cytokine-175 Magnetic Bead Array Kit

This RayPlex® kit is a cytometric bead array (CBA) that uses magnetic beads to quantitatively measure, by flow cytometry, 175 key human cytokine biomarkers. The assay has been optimized for serum and cell culture supernatant.

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Catalog #:
FAHM-CYT-175
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Product Description

Specifications

Size 8 samples, 16 samples, 40 samples
Species Human
Quantitative/Semi-Quantitative Quantitative
Number of Targets Detected 175
Specificity No detectable or significant cross reactivity was detected between antibodies or targets.
Compatible Sample Types Cell Culture Supernatants, Serum
Solid Support Fluorescent Microbeads
Method Of Detection Flow Cytometry
Design Principle Sandwich-based
Estimated Lead Time 1-2 business days
Shipping Type Blue ice
Storage 4°C
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Introduction

Soluble proteins play vital roles in homeostasis and disease development, including innate immunity, apoptosis, angiogenesis, cell growth, and differentiation. Thus, protein profiling has research and clinical applications, such as disease biomarker discovery and drug development. The traditional technique for detecting and quantifying soluble proteins is the enzyme-linked immunosorbent assay (ELISA), which only allows detection of one analyte each time. However, using this method to detect multiple proteins – particularly for precious samples with small quantities – becomes challenging as the processing time and required sample volume increase.

RayPlex® is a multiplexed sandwich- and bead-based quantitative antibody array for the simultaneous detection of multiple proteins using small sample volumes. It couples the versatility of RayBiotech's vast antibody pair library with familiar, reliable flow cytometry methodology. Together this creates a multiplex bead-based array requiring no dedicated instrumentation, only common flow cytometers equipped with blue (or green) and red lasers.

The RayPlex® Human Cytokine-175 Magnetic Bead Array Kit is designed to assay 175 common human cytokines simultaneously. The kit uses magnetic beads that allow the use of an automatic plate washing system with magnetic separation ability during washing steps. For data in this manual, a BioTek 405 TS Washer and LifeSep™ 96F magnetic separator unit were used. Further details are included in Section VI.

Product Features

  • Less sample, more data: up to 25 proteins can be quantified from only 25 µl or less of your precious sample
  • 4 hour processing time
  • 5-10x more cost-effective than ELISA
  • Compatible with most flow cytometers
  • No specialized or dedicated instrument(s) needed
  • High specificity and system reproducibility
  • Well-suited for high throughput assays

Target Names

Eotaxin-2 / MPIF-2 / CCL24, BLC / BCA-1 / CXCL13, RANTES / CCL5, IL-10 / IL10, MCP-1 / CCL2, PDGF-BB / PDGFB, IFN-gamma / IFNG, TNF alpha / TNFA, IL-5 / IL5, IL-1 beta / IL-1F2 / IL1B, IL-6 / IL6, IL-2 / IL2, IL-4 / IL4, GCSF / CSF3, IL-12 p70 / IL12A, IL-8 / IL8 / CXCL8, MIP-1 alpha / CCL3, IL-7 / IL7, IL-9 / IL9, TIMP-2 / TIMP2, TNF R1 / TNFRSF1A, IL-1Ra / IL-1RN / IL-1 F3 / IRAP / IL1RN, TNF R2 / TNFRSF1B, IL-17A / IL17A, I-309 / TCA-3 / CCL1, IL-12 p40 / IL12B, IL-13 / IL13, IL-15 / IL15, Eotaxin / CCL11, IL-16 / CXCR4 / IL16, MIP-1 delta / CCL15, TNF beta / TNFSF1 / LT-alpha / TNFB, IL-1 alpha / IL-1F1 / IL1A, ICAM-1 / CD54 / ICAM1, IL-11 / IL11, IFN-alpha / IFNA1, M-CSF / CSF1, Common beta Chain / GM-CSF R beta / CSF2RB / GM-CSF, IL-28A / IFN lambda 2 / IL28A, MIP-3 beta / CCL19, MIG / CXCL9, TSLP, Eotaxin-3 / CCL26, MIP-1 beta / CCL4, MDC / CCL22, I-TAC / CXCL11, TECK / CCL25, CCL16 / HCC-4, IL-18BPa / IL18BP, NAP-2 / TCK-1 / CXCL7 / PPBP, Osteopontin / SPP1, MIF, LIF, Axl / UFO, Betacellulin / BTC, ENA-78 / CXCL5 / LIX, Lymphotactin / XCL1, SDF-1 alpha / CXCL12 alpha, CK beta 8-1 / CCL23 / MPIF-1, CXCL1 / GRO alpha / KC / CINC-1, PARC / CCL18, IP-10 / CXCL10, MIP-3 alpha / CCL20, TARC / CCL17 / ABCD-2, MCP-2 / CCL8, MCP-3 / MARC / CCL7, GCP-2 / CXCL6, CCL14 / HCC-1 / HCC-3, IL-31 / IL-31R alpha / IL31RA, IL-29 / IL29 / IFN-lambda 1 / IL-10R beta / IL10RB, IL-17F / IL17F, MCP-4 / CCL13, MEC / CCL28, CTACK / CCL27, AgRP, Angiostatin / Plasminogen / PLG, LAP / TGF beta 1 / TGFB1, IL-2R alpha / IL2RA, Angiogenin / ANG, PDGF-AB, E-Cadherin / CDH1, Activin A / INHBA, Cripto-1 / TDGF1, ICAM-2 / CD102 / ICAM2, IL-2R beta / CD122 / IL2RB, VEGFR1 / FLT1, FLRG / Follistatin / FST, Cathepsin S / CTSS, CD40 / TNFRSF5, Fc gamma RIIB/C / CD32B/C / FCGR2B/C, gp130 / sgp130 / IL6ST, Thrombopoietin / TPO / THPO, DAN / PARN, Galectin-7 / LGALS7, Fas Ligand / TNFSF6 / CD178 / FASLG, SDF-1 beta / CXCL12 beta, VEGFC, TGF beta 2 / TGFB2, IL-13R alpha 1 / IL13RA1, IL-1R-4 / ST2 / IL-33R / IL1RL1, Tie-2 / TEK / TIE2, TSH / TSHB, Sonic Hedgehog N-Terminal / Shh-N, TREM-1 / TREM1, IL-23 / IL23A / IL-23 p19, TRAIL R4 / TNFRSF10D, Alpha-Fetoprotein / AFP, Angiopoietin-like protein 4 / ANGPTL4, CA125 / MUC16, FSH / FSHB, hCG beta / CGB3, IGF-1 R / IGF1R, IL-1R-2 / IL-1RT-2 / IL-1R beta / CD121b / IL1R2, IL-18R beta / IL-1R-7 / AcPL / IL-18RAcP / IL18RAP, IL-21 / IL21, IL-3 / IL3, Leptin / Lenep, MMP-10 / MMP10, MMP-13 / MMP13 / Pro-MMP-13, MMP-3 / MMP3, MMP-8 / MMP8, Enolase 2 / ENO2 / NSE, Oncostatin M / OSM, Prolactin / PRL, PSA-free / KLK3, Siglec-9 / CD329 / SIGLEC9, ADAM-17 / TACE / ADAM17, TIMP-4 / TIMP4, TRAIL R3 / TNFRSF10C, DR6 / TNFRSF21, CD14, CEACAM1, IL-1R-1 / IL-1RT-1 / IL-1R alpha / CD121a / p80 / IL1R1, PECAM-1 / CD31 / PECAM1, E-Selectin, Endoglin / CD105 / ENG, Trappin-2 / PI3, ErbB3, CD30 / TNFRSF8, IL-2R gamma / Common gamma Chain / IL2RG, CD80 / B7-1, Contactin-2 / TAG1 / CNTN2, uPAR / PLAUR, Dtk / TYRO3, ALCAM / CD166, LIMPII / LIMP2 / SCARB2, MICA, TIM-1 / KIM-1 / HAVCR1, IL-17RA / IL17RA, XEDAR / EDA2R, Fas / Apo-1 antigen / TNFRSF6 / CD95, IL-21R / IL21R, CD40 Ligand / TNFSF5 / CD40LG, Flt-3 Ligand / FLT3L, GITR / TNFRSF18, GDF-15 / GDF15, Osteoprotegerin / TNFRSF11B / OPG, VEGFD / FIGF, beta-NGF, Growth Hormone / GH1, IGF-1 / IGF1, NGFR / TNFRSF16, TGF beta 3 / TGFB3, BMP-4 / BMP4, IGFBP-2 / IGFBP2, BMP-7 / BMP7, VEGFA, bFGF / FGF-2 / FGF2, LAP / TGF beta 1 / TGFB1, SCF / KITLG, IGFBP-6 / IGFBP6, VEGFR3 / FLT4, KDR / VEGFR2, PIGF, EG-VEGF / PK1 / PROK1

Application Notes

Kit Components
  • RayPlex™ Multiplex Bead Cocktail
  • 20X Sample Dilution Buffer
  • 1X Assay Diluent
  • 20X Wash Buffer
  • RayBio® Serum Diluent
  • Lyophilized Protein Standard Mix
  • Biotinylated Detection Antibody Cocktail
  • 10X Streptavidin-PE
  • Flow Cytometer Setup Bead Cocktail
  • Flat-bottom 96-well Microplate
  • Filter 96-well Microplate
Other Materials Required
  • Orbital 96-well plate shaker (with ability to reach 1000 rpm)
  • Flow Cytometer with blue (or green) and red lasers capable of measuring phycoerythrin (PE), Alexa Fluor 700 (AF700), and Alexa Fluor 647 (AF647)/APC channels
  • Aluminum foil or 96-well Plate Aluminum Sealers
  • Deionized (DI) water
  • 1.5 ml polypropylene microcentrifuge tubes or similar
  • Microcentrifuge
  • Recommended: Automated plate washing system with magnetic separation
  • Optional: 96-well round bottom plate (to prepare samples)
  • Optional: High throughput sampler (HTS) for 96-well plate reading
  • Optional: Vacuum manifold compatible with a 96-well microplate if using a Filter 96-Well Microplate

Don't have a compatible flow cytometer? RayBiotech provides full testing services for RayPlex™ bead arrays. Get a quote!
Protocol Outline
  1. Prepare Reagents, Standards, and Samples
  2. Add RayPlex™ Multiplex Bead Cocktail to each well
  3. Incubate with Samples and Standards
  4. Incubate with Biotinylated Detection Antibody Cocktail
  5. Incubate with Streptavidin-Conjugated Fluorophore
  6. Analyze with a compatible flow cytometer
  7. Perform data analysis

Storage/Stability

Upon receipt, the Lyophilized Protein Standard Mix should be stored at -20°C or below. Other components should be stored at 4°C. Use reagents immediately after preparation. Additional vials of the Lyophilized Protein Standard Mix can be purchased separately. If stored at the proper temperature, the kit will retain complete activity for at least 6 months.
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