Mab206
Manufactured by R&D Systems
Sourced in United States
MAB206 is a monoclonal antibody product developed by R&D Systems. It recognizes and binds to a specific target antigen. The core function of this product is to serve as a research tool for the detection and study of the target antigen in various applications.
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Lab products found in correlation
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Mab250, by R&D Systems (1 mentions)
Mab218, by R&D Systems (1 mentions)
N hydroxysuccinimide, by Merck Group (1 mentions)
1 ethyl 3 3 dimethylaminopropyl carbodiimide, by Merck Group (1 mentions)
Mab601, by R&D Systems (1 mentions)
Baf210, by R&D Systems (1 mentions)
Tween 20, by Thermo Fisher Scientific (1 mentions)
Igg1 isotype control, by R&D Systems (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
Dimethylformamide, by Thermo Fisher Scientific (1 mentions)
Triethylamine, by Thermo Fisher Scientific (1 mentions)
Cysteamine, by Merck Group (1 mentions)
Succinic anhydride, by Merck Group (1 mentions)
16 mercaptohexadecanoic acid, by Merck Group (1 mentions)
Mab610, by R&D Systems (1 mentions)
Clone 6708, by R&D Systems (1 mentions)
Clone 11711, by R&D Systems (1 mentions)
12 protocols using mab206
1
Cytokine Neutralization in Cell Culture
2
Multiplex Cytokine Sandwich Immunoassay
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The following reagents were from Sigma–Aldrich (St. Louis, MO, USA); 16-mercaptohexadecanoic acid, cysteamine, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, succinic anhydride and N-hydroxysuccinimide. Triethylamine, dimethylformamide and bovine serum albumin (BSA) were from Fisher Scientific (Pittsburgh, PA, USA). Six-armed-poly(ethyleneglycol)–amine was from SunBio (South Korea). PBS (Cat#70011-036) and Tween-20 were from Life Technologies (Grand Island, NY, USA).
Sandwich antibody pairs for IL-20 were produced by Novo Nordisk A/S. Capture anti-IL-20 antibody was of the IgG1 isotype. Unspecific binding was assessed using a IgG1 isotype control from R&D Systems (MAB002, Minneapolis, MN, USA). IRDye-800 streptavidin conjugate was from LiCor Biosciences (Lincoln, NE, USA). Mouse, goat and bovine IgG were from Jackson ImmunoResearch (West Grove, PA, USA).
Purified cytokine antigen standards and sandwich antibody pairs for IL-1β (MAB601, BAF201), IL-10 (MAB2172, BAF217), IL-6 (MAB206, BAF206), and TNFα (MAB610, BAF210) were purchased from R&D systems. All capture antibodies were of the IgG1 isotype.
Sandwich antibody pairs for IL-20 were produced by Novo Nordisk A/S. Capture anti-IL-20 antibody was of the IgG1 isotype. Unspecific binding was assessed using a IgG1 isotype control from R&D Systems (MAB002, Minneapolis, MN, USA). IRDye-800 streptavidin conjugate was from LiCor Biosciences (Lincoln, NE, USA). Mouse, goat and bovine IgG were from Jackson ImmunoResearch (West Grove, PA, USA).
Purified cytokine antigen standards and sandwich antibody pairs for IL-1β (MAB601, BAF201), IL-10 (MAB2172, BAF217), IL-6 (MAB206, BAF206), and TNFα (MAB610, BAF210) were purchased from R&D systems. All capture antibodies were of the IgG1 isotype.
3
Comparative Migration and Invasion Assay
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Approximately 50 000 cells/insert of parental/BRAFi‐R HTB63, A375, and A2058 cells were used to compare their migratory and invasive capacities. In separate experiments, appropriate controls [NaHCO3 buffer, pH ~7 for Box5 (WNT5A antagonisitic peptide, Jenei et al., 2009 ) treatment, IgG1 isotype antibody (R&D Systems, Minneapolis, MN, USA, Clone #11711) for IL‐6 Ab (R&D Systems, Clone #6708, MAB206) exposure, DMSO for ML141 treatments, 48 h] were used to compare the Box5 (200 μm ) and/or IL‐6 Ab (2 μg·mL−1) and ML141 (2.5 μm ) treatment effects on the cell migration and invasion of HTB63‐R and A375‐R cells. After each experiment, photographs of cells in the inserts were captured using an inverted light microscope (Nikon TMS), and the numbers of migrated/invaded cells were counted using nih imagej ® (NIH Image J, Bethesda, MD, USA) software. The results are presented as the relative migration/invasion compared with parental or vehicle‐treated cells.
4
Immunofluorescence analysis of MMP9 and IL-6 in human corticotroph adenomas
5
Sphere Formation Assay for Cancer
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To generate spheres in vitro, we resuspended ~1.0 × 106 the parental A549 or ~1.0 × 106 OSK-A549-colony cells, ~3.0 × 105 HUVECs, and ~3.0 × 105 MSCs were resuspended in sphere forming medium and seeded on a low-attachment 24-well flat plate (Prime surface Sumitomo). We adopted the following specific ratios with reference to a previous report24 (link). A549 or OSK-A549-Colony cells: HUVEC: MSC = 10:1:4 ~ 5:4:4. During 6 to 12 days of culture, the self-organized spheres were photographed using a BZ8000 microscope (Keyence) and were pathologically analyzed. To assess the chemosensitivity of the spheres and the IL-6 function of the forming spheres, the spheres were co-cultured with or without 5 μM of cisplatin, 1 μg/ml of anti-IL-6 antibody (R & D, MAB206) and 10 ng/ml of IL-6 protein from the first day of this assay. The photographed pictures were then analyzed to calculate fluorescence intensity using the ImageJ software program.
6
Quantification of Immune Mediators in Cell Culture
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The levels of PGE2 (#514010, Cayman Chemical, Ann Arbor, MI, USA), periostin (#DY3548B, R&D systems), LIF (#DY7734-05, R&D systems), IFN-γ (#3420-1H-6, Mabtech), and Granzyme B (#3585-1H-6, Mabtech) in cell culture supernatants were measured with ELISA Kits according to manufacturer´s instructions. For IL-6, IL-10 and IL-17 an in-house ELISA was set up, using a monoclonal anti-human capture antibody #MAB206, #MAB127107 and #MAB317 (R&D Systems), a standard of the recombinant human antibody #206-IL, #1064-IL, #317IL (R&D systems), and a biotinylated anti-human antibody #BAF206, #BAF217, #BAF317 (R&D systems), accordingly. A streptavidin conjugated to poly-horseradish peroxidase was used for all the enzymatic reaction (#M2032, Sanquin, Amsterdam, Netherlands).
7
IL-6 ELISA for Chondrocyte Cultures
8
Cytokine Profiling in Blood and Cells
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The concentration of cytokines in peripheral blood and cell culture supernatants was analyzed as previously described (43 (link), 44 (link)). Briefly, cytokines in the plasma of peripheral blood of children and cytokines released by non-stimulated and stimulated DCs after 24 h of stimulation with EcO83 or lipopolysaccharide (LPS) (1 µg/ml, cat. no. L2654-1MG; Sigma-Aldrich, St. Louis, MO, USA) were detected by ELISA. Reagents for IL-4, IL-5, IL-6, IL-10, IL-13, interferon (IFN)-γ, and transforming growth factor (TGF)-β detection were purchased from R&D Systems (Minneapolis, MN, USA) (IL-4: primary antibody MAB 604, biotinylated secondary antibody BAF 204, recombinant standard protein 204-IL; IL-5: MAB 405, BAM 6051, 205-IL; IL-6: MAB 206, BAF 206, 206-IL; IL10: MAB 2172, BAF 2017, 217-IL; IL-13: MAB 213, BAF 285, 285-IL; IFN-γ: MAB 2852, BAF 285, 285-IF; TGF-β: MAB 240, BAF 240, 240-B). Concentration of IL-10 in cell culture supernatants was determined by DUO SET DY217B (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.
9
Boyden Chamber Cell Migration Assay
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A Boyden chamber migration assay was used to quantify the number of migratory cells (Costar; Corning Incorporated, NY, USA). ACP cells that had been serum starved for 6 h were resuspended in serum-free culture (1×105 cells/300 µl) medium containing IL-6 (0–100 ng/ml) or IL-6 (100 ng/ml)/anti-hIL-6- antibody (10 µg/ml; MAB206; R&D Systems, Inc.) and placed in the top chamber of the inserts (1×105 cells/well). Following incubation for 20 h (37°C, 5% CO2), the inserts were removed; cells were fixed and stained with Giemsa solution (Solarbio Life Sciences, Beijing, China) at room temperature for 20 min, and were viewed and counted under a light microscope. Images of representative microscopic fields were captured. Quantification of cell migration was expressed as the mean count of stained cells in 5 random fields of each filter. Each experiment was repeated three times (n=3).
10
IL-6 Cytokine Regulation of MLS Cell Lines
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MLS cell lines 402-91, 1765-92, and 2645-94 were seeded to 96 well microtiter plates at 2000 cells/well and allowed to settle for 8 hours. The culture medium was replaced with medium supplemented with 3% fetal bovine serum, with and without 1 unit/mL of recombinant IL6 and with or without 0.5 μg/mL of monoclonal IL6 antibody mAB 206 (R&D systems). Trypan blue excluding cells were counted in an inverted microscope after 48 hours of incubation. An Epstein-Barr virus immortalized lymphoblastoid cell line was included as positively responding control [20 (link)].
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