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Bio plex pro human chemokine panel 40 plex

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The Bio-Plex Pro™ Human Chemokine Panel 40-Plex is a multiplex assay designed to simultaneously detect and quantify the levels of 40 different human chemokines in a single sample. The panel utilizes bead-based technology and can be analyzed using a Bio-Plex® or similar flow cytometry-based platform.

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Lab products found in correlation

Bio plex 200 system, by Bio-Rad (6 mentions) Envision flex target retrieval solution, by Agilent Technologies (2 mentions) Hcg human procartaplex simplex kit, by Thermo Fisher Scientific (2 mentions) Bio plex pro human inflammation panel 1 il 28a ifn λ2, by Bio-Rad (2 mentions) Bio plex wash station, by Bio-Rad (2 mentions) Bio plex magpix multiplex reader, by Bio-Rad (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Cd8 percp clone sk1, by BD (1 mentions) Envision g 2 system ap rabbit mouse, by Agilent Technologies (1 mentions) Envision duoflex doublestain system, by Agilent Technologies (1 mentions) One step reverse transcription kit, by Qiagen (1 mentions) Precellys 24 homogenizer, by Roche (1 mentions) Easypack protease inhibitor cocktail, by Roche (1 mentions) Human ifn γ, by Thermo Fisher Scientific (1 mentions) Tumor necrosis factor (tnf), by Thermo Fisher Scientific (1 mentions) Ccl20, by Thermo Fisher Scientific (1 mentions) Cxcl10, by Thermo Fisher Scientific (1 mentions) Cxcl8, by Thermo Fisher Scientific (1 mentions) Human magnetic luminex assay, by R&D Systems (1 mentions) Lsrfortessa cell analyzer, by BD (1 mentions)

29 protocols using bio plex pro human chemokine panel 40 plex

1

Multiplex Analysis of Immune Markers

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Materials were from Sigma Aldrich (St. Louis, Mo, USA) except rK39 immunochromatographic test strips (InBios International, Seattle, WA, USA), anti-human CD4 (clone 4B12, Novocastra, Newcastle upon Tyne, UK,), Chemokine Receptor 4 (CCR4, clone 1G1), CD3 Peridinin chlorophyll (PerCP, clone SK7), CD8 Alexa Fluor 488 (clone RPA-T8), CD8 PerCP (clone SK1), CCR4 Phycoerythrin (PE, clone 1G1, BD Biosciences, San Jose, CA, USA), PD-1(clone EH12.2H7) and CD127 (clone A019D5) APC (Biolegend, San Diego, CA, USA), IL-5 (clone 9906, R&D Systems, Minneapolis, MN, USA), CCL17 (clone N-20), IL-10 (clone E-10), PD-1 (clone C-20), Perforin H (clone H-315), Granzyme B (clone 2C5, Santa Cruz Biotechnology, Dallas, TX, USA), antibody diluent, Target Retrieval Solution Citrate pH 6 (S2369), secondary detection system EnVision™ G|2 System/AP-Rabbit/Mouse (Permanent Red), EnVision™ FLEX Target Retrieval Solution, EnVision™ DuoFLEX Doublestain System (Dako, Glostrup, Denmark), RNALater and RNAqueous kit (Ambion, Austin, TX, USA), One-step reverse transcription kit from Qiagen (Hilden, Germany) and Bio-Plex Pro™ Human Chemokine Panel 40-Plex (BioRad, Hercules, CA, USA).
Mukherjee S., Sengupta R., Mukhopadhyay D., Braun C., Mitra S., Roy S., Kanti Das N., Chatterjee U., von Stebut E, & Chatterjee M. (2019). Impaired activation of lesional CD8+ T-cells is associated with enhanced expression of Programmed Death-1 in Indian Post Kala-azar Dermal Leishmaniasis. Scientific Reports, 9, 762.
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2

Cytokine/Chemokine Profiling of Tumor Xenografts

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Cytokine/chemokine analyses from tumors of humanized mice were conducted using the Bio-Plex Pro™ Human Chemokine Panel, 40-Plex (Bio-Rad Laboratories AG, Cressier, Switzerland, Cat No.: 171AK99MR2). Small tumor fragments were snap frozen and whole protein was isolated in the presence of EASYpack Protease Inhibitor Cocktail (Roche; ref 5892970001) using the Precellys®24 Homogenizer and Bio-Plex® Cell Lysis Buffer following manufacturer instructions. Whole protein content was measured with BCA™ Protein Assay Kit (Thermo Scientific) before cytokine measurement was performed.
Sam J., Colombetti S., Fauti T., Roller A., Biehl M., Fahrni L., Nicolini V., Perro M., Nayak T., Bommer E., Schoenle A., Karagianni M., Le Clech M., Steinhoff N., Klein C., Umaña P, & Bacac M. (2020). Combination of T-Cell Bispecific Antibodies With PD-L1 Checkpoint Inhibition Elicits Superior Anti-Tumor Activity. Frontiers in Oncology, 10, 575737.
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3

Multiplex Cytokine Profiling of 3D Spheroids

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The concentration of 43 soluble factors in the supernatant was determined by Luminex multiplex technology. The supernatant was harvested at day 14 from spheroids with and without monocytes added on day 7. The following kits were purchased from Bio-Rad Laboratories, Inc. (Hercules, CA, USA): Bio-Plex Pro™ Human Chemokine Panel, 40-Plex (#171AK99MR2), Bio-Plex Pro Human IL-1ra Set (#171B5002M), Bio-Plex Pro Human IL-12 (p70) Set (#171B5011M) and Bio-Plex Pro™ Human VEGF Set (#171B5027M). All kits were used according to the manufacturer’s instructions. Median fluorescence intensity (MFI) was read on a Luminex MAGPIX® reader with XPONENT software.
Madsen N.H., Nielsen B.S., Nhat S.L., Skov S., Gad M, & Larsen J. (2021). Monocyte Infiltration and Differentiation in 3D Multicellular Spheroid Cancer Models. Pathogens, 10(8), 969.
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4

Quantifying Cytokine Modulation by ISG15

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CircuLex Human ISG15 ELISA Kit [#CY-8085, Nordic Biosite]; human IFNγ [#88-7316-86, Invitrogen, Thermo Fisher Scientific, Bremen, Germany]; TNF [#DY210], CXCL10 [#DY266], CCL20 [#DY360] and CXCL8 [#DY208]; all [R&D]; were used according to the manufacturers’ protocol. Cytokines in conditioned medium from stimulated PBMCs were detected and analysed by Bio-Plex Pro™ Human Chemokine Panel, 40-Plex [#171ak99mr2, Bio-Rad laboratories] according to the manufacturer’s protocol. The Limulus Amebocyte Lysate [LAL]-assay [#50-647U, LONZA Walkersville, MD USA, QCL-100TM] was performed according to the manufacturer’s instructions to confirm that the levels of endotoxin were well below 1 EU [<0.1 ng] per µg protein, as stated by the suppliers of both recombinant IL12 and ISG15. Nevertheless, since even 0.02 ng/ml LPS has been shown to significantly stimulate small amounts of cytokine production,28 (link) we defined cytokine release to be modulated by ISG15 only if the enhanced cytokine concentration in conditioned medium was above 1000 pg/ml compared with untreated control in the multiplex assay.
Østvik A.E., Svendsen T.D., Granlund A.V., Doseth B., Skovdahl H.K., Bakke I., Thorsvik S., Afroz W., Walaas G.A., Mollnes T.E., Gustafsson B.I., Sandvik A.K, & Bruland T. (2020). Intestinal Epithelial Cells Express Immunomodulatory ISG15 During Active Ulcerative Colitis and Crohn’s Disease. Journal of Crohn's & Colitis, 14(7), 920-934.
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5

Multiplex Cytokine Measurement

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Plasma cytokines levels (TNF-α, IL-6, IL-1β and MCP/CCL2) were measured using the Bio-Plex Pro™ Human Chemokine Panel, 40-plex (Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer's instruction.
Zhou Z., Guille C., Ogunrinde E., Liu R., Luo Z., Powell A, & Jiang W. (2017). Increased systemic microbial translocation is associated with depression during early pregnancy. Journal of psychiatric research, 97, 54-57.
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6

Multiplex Chemokine Profiling of Cell Media

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The culture medium derived from U251‐sgNC, sg2, and sg3 cells was analyzed for the presence of 40 chemokines using the commercial Bio‐Plex Pro™ Human Chemokine Panel 40‐Plex (Bio‐Rad Laboratories, Inc., CA, United States), following the manufacturer's instructions. A volume of 50 μL of the undiluted sample was subjected to the assay.
Huang L., Wang Z., Liao C., Zhao Z., Gao H., Huang R., Chen J., Wu F., Zeng F., Zhang Y., Jiang T, & Hu H. (2024). PVT1 promotes proliferation and macrophage immunosuppressive polarization through STAT1 and CX3CL1 regulation in glioblastoma multiforme. CNS Neuroscience & Therapeutics, 30(1), e14566.
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7

Comprehensive Chemokine Profiling in Serum Samples

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Serum samples were prepared by centrifugation at 300×g for 15 min and stored at −80°C. A Bio-Plex Pro™ Human Chemokine Panel (40 plex) (Bio-Rad, Hercules, CA) was used in accordance with the manufacturer’s instructions. The complete panel screened for expression of the following chemokines: CCL1, CCL2, CCL3, CCL7, CCL8, CCL11, CCL13, CCL15, CCL17, CCL19, CCL20, CCL21, CCL22, CCL23, CCL24, CCL25, CCL26, CCL27, CX3CL1, CXCL1, CXCL2, CXCL5, CXCL6, CXCL8, CXCL9, CXCL10, CXCL11, CXCL12, CXCL13, CXCL16, GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-16, MIF, and TNF-α.
Sakata T., Yoshio S., Yamazoe T., Mori T., Kakazu E., Aoki Y., Aoyanagi N., Okamoto T., Ito T., Toyoda H., Kawaguchi T., Ono Y., Takahashi Y., Taketomi A, & Kanto T. (2024). Immunoglobulin-like transcript 2 as an impaired anti-tumor cytotoxicity marker of natural killer cells in patients with hepatocellular carcinoma. Frontiers in Immunology, 15, 1389411.
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8

Multiplex Luminex Assays for Placental Proteins

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Luminex assays were performed using the following kits according to the manufacturer’s instructions: hCG Human ProcartaPlex Simplex Kit (Invitrogen, EPX010-12388-901), Bio-Plex Pro Human Inflammation Panel 1 IL-28A / IFN-λ2 (Bio-rad, 171BL022M), Bio-Plex Pro Human Inflammation Panel 1, 37-Plex (Bio-rad, 171AL001M), and Bio-Plex Pro Human Chemokine Panel, 40-Plex (Bio-rad, 171AK99MR2). Plates were washed using the Bio-Plex wash station (Bio-rad, 30034376) and read on a Bio-Plex 200 system (Bio-rad, 171000205). All samples from both polarity conditions (STBin and STBout) were tested in duplicate, and each condition was performed with at least three biological replicates from unique placental tissue. All measurements were normalized to total protein of either STBin or STBout TOs wells, quantified as described above.
Yang L., Liang P., Yang H, & Coyne C.B. (2023). Trophoblast organoids with physiological polarity model placental structure and function. bioRxiv.
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9

Multiplex ELISA Profiling of Secreted Factors

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Supernatants from the BMSC samples were evaluated using a multiplex ELISA method. In total, 40 soluble factors were measured using the Bio-Plex Pro™ Human Chemokine Panel, 40-plex (Bio-Rad Laboratories, Hercules, CA, USA) and a Bio-Plex® MAGPIX™ Multiplex Reader (Bio-Rad Laboratories), according to the manufacturer’s instructions. In brief, supernatant samples were incubated with microbeads coated with antibodies specific to the above-mentioned chemokines for 1 hour. After washing, the beads were incubated with the detection antibody cocktail, which included a biotinylated antibody specific to each single cytokine. After another wash step, the beads were incubated with streptavidin-phycoerythrin for 10 minutes and washed again. Finally, the fluorescence was measured using a Bio-Plex® MAGPIX™ Multiplex Reader. Partek Genomic Suite 6.4 (Partek Inc., St. Louis, MO, USA) was used for data visualization and hierarchical cluster analysis. The data was normalized using log(X + offset).
Jin P., Zhao Y., Liu H., Chen J., Ren J., Jin J., Bedognetti D., Liu S., Wang E., Marincola F, & Stroncek D. (2016). Interferon-γ and Tumor Necrosis Factor-α Polarize Bone Marrow Stromal Cells Uniformly to a Th1 Phenotype. Scientific Reports, 6, 26345.
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10

Multicolor Flow Cytometry Analysis of Immune Cells

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To evaluate the phenotypes of immune cells, a multicolor flow‐cytometric analysis was performed with an LSR Fortessa cell analyzer (BD, San Jose, CA) and the FlowJo software (Tree Star, Inc., Ashland, OR). The cells analyzed and the gating strategies are shown in Supporting Fig. S1. The concentrations of cytokines and chemokines in serum samples were examined with the Bio‐Plex Pro Human Chemokine Panel, 40‐Plex (Bio‐Rad, Hercules, CA) and a customized Human Magnetic Luminex Assay (R&D Systems, Minneapolis, MN) on the Bio‐Plex 3D Suspension Array System. The reagents are listed in Supporting Table S3.
Doi H., Yoshio S., Yoneyama K., Kawai H., Sakamoto Y., Shimagaki T., Aoki Y., Osawa Y., Yoshida H, & Kanto T. (2019). Immune Determinants in the Acquisition and Maintenance of Antibody to Hepatitis B Surface Antigen in Adults After First‐Time Hepatitis B Vaccination. Hepatology Communications, 3(6), 812-824.
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