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Tnfα mab11

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TNFα (MAb11) is a monoclonal antibody directed against the human tumor necrosis factor alpha (TNFα) protein. It is a research-use-only laboratory reagent.

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Lab products found in correlation

Il 2 mq1 17h12, by BioLegend (2 mentions) Pd 1 j105, by Thermo Fisher Scientific (2 mentions) Cd127 ebiordr5 and cd218a h44, by Thermo Fisher Scientific (2 mentions) Nkp44 2.9, by Miltenyi Biotec (2 mentions) Cd28 cd28.2, by Beckman Coulter (2 mentions) Cd62l dreg 56, by Thermo Fisher Scientific (1 mentions) Dnam 1, by BioLegend (1 mentions) Ulbp 1 170818, by R&D Systems (1 mentions) Cytofix cytoperm kit, by BD (1 mentions) Cd107a pe h4a3, by BD (1 mentions) Brefeldin a, by BD (1 mentions) Monensin, by BD (1 mentions)

4 protocols using tnfα mab11

1

Polychromatic Cytometry and Cell Sorting

Cited 1 time
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Polychromatic flow cytometry and cell sorting were performed on stained mononuclear cells as previously described15 (link). Antibodies against the following antigens were used for staining at predetermined concentrations: CD4 (clone OKT4), CD8 (SK1), IFNγ (4S.B3), IL-17 (eBio64DEC17), IL-21 (3A3-N2), IL-22 (IL22JOP), TNFα (MAb11), and PD-1 (J105) from eBioscience; CCR5 (3A9), CD3 (SP34-2), CD8 (SK1), CD16 (HI149), CD20 (2H7), CD23 (M-L233), CD45 (D058-1283), CD123 (7G3), and HLA-DR (L243) from BD; c-Kit (104D2), CD1a (HI149), CD11c (3.9), CD14 (M5E2), CD34 (561), CD95 (DX2), and IL-2 (MQ1-17H12) from Biolegend; CD127 (eBioRDR5) and CD218a (H44) from Thermo; NKp44 (2.9) from Miltenyi; and CD28 (CD28.2) from Beckman Coulter. CD4+ and CD8+ TM were defined as CD95+ singlet, clean, live, CD3+ lymphocytes. NKp44+ ILC3s were defined as CD45+ singlet, clean, live, lineage-negative (CD1a, CD3, CD8, CD11c, CD14, CD16, CD20, CD23, CD34, CD123), NKp44+CD127+CD218+ lymphocytes. Positive/negative gating based on clearly grouped populations, historical-determined expression, and the use of internal controls (Supplementary Fig. 8). A threshold of 100 collected events in the parent population was utilized for all subset expression analysis.
Ortiz A.M., Flynn J.K., DiNapoli S.R., Vujkovic-Cvijin I., Starke C.E., Lai S.H., Long M.E., Sortino O., Vinton C.L., Mudd J.C., Johnston L., Busman-Sahay K., Belkaid Y., Estes J.D, & Brenchley J.M. (2018). Experimental Microbial Dysbiosis Does Not Promote Disease Progression in SIV-Infected Macaques. Nature medicine, 24(9), 1313-1316.
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2

Phenotyping of Expanded Natural Killer Cells

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The phenotype of eNK cells was analyzed weekly using flow cytometry. The following mouse anti-human antibodies were used for staining: CD16 (3G8), CD56 (NCAM 16.2), NKG2D (1D11), DNAM-1 (DX11), CD3 (HIT3a) from BD PharMingen; CXCR1 (8F1), CXCR3 (CEW33D), CXCR4 (L276F12), CXCR6 (K041E5), CCR4 (L291H4), CCR5 (J418F1), CCR6 (G034E3), CCR9 (L053E8), CX3CR1 (2A9-1), Fas-L (NOK-1), IFNγ (B27) from BioLegend; CD62L (DREG-56), intracellular Perforin (dG9), Granzyme B (GB11), and TNF-α (MAb11) from eBioscience. For surface staining, NK cells were resuspended in FACS buffer (0.5% FBS and 2 mM EDTA in PBS), blocked with 10% mouse serum for 30 min, then incubated with the indicated antibodies for 30 min at 4°C using concentrations recommended by the manufacturers. For subsequent intracellular staining, NK cells were stained using the BD Cytofix/Cytoperm Kit following manufacturer protocol. Data was acquired using a BD FACS Accuri C6 and analyzed using FlowJo v10.2 software. Isotype IgG antibodies were used as negative controls. Human NK cells were defined as CD16+, CD56+ and CD3−. Mouse anti-human HLA-ABC (W6/32), MICA/B (6D4), CD155 (2H7) and PD-L1 (MIH1) from eBioscience, ULBP-1 (170818) from R&D, and Fas (DX2) from BioLegend were used to determine MHC-I, NKG2D ligand and DNAM-1 ligand expression on TC106 with surface staining performed as above for NK cells.
Tong A.A., Hashem H., Eid S., Allen F., Kingsley D, & Huang A.Y. (2017). Adoptive natural killer cell therapy is effective in reducing pulmonary metastasis of Ewing sarcoma. Oncoimmunology, 6(4), e1303586.
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3

Polychromatic Cytometry and Cell Sorting

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Polychromatic flow cytometry and cell sorting were performed on stained mononuclear cells as previously described15 (link). Antibodies against the following antigens were used for staining at predetermined concentrations: CD4 (clone OKT4), CD8 (SK1), IFNγ (4S.B3), IL-17 (eBio64DEC17), IL-21 (3A3-N2), IL-22 (IL22JOP), TNFα (MAb11), and PD-1 (J105) from eBioscience; CCR5 (3A9), CD3 (SP34-2), CD8 (SK1), CD16 (HI149), CD20 (2H7), CD23 (M-L233), CD45 (D058-1283), CD123 (7G3), and HLA-DR (L243) from BD; c-Kit (104D2), CD1a (HI149), CD11c (3.9), CD14 (M5E2), CD34 (561), CD95 (DX2), and IL-2 (MQ1-17H12) from Biolegend; CD127 (eBioRDR5) and CD218a (H44) from Thermo; NKp44 (2.9) from Miltenyi; and CD28 (CD28.2) from Beckman Coulter. CD4+ and CD8+ TM were defined as CD95+ singlet, clean, live, CD3+ lymphocytes. NKp44+ ILC3s were defined as CD45+ singlet, clean, live, lineage-negative (CD1a, CD3, CD8, CD11c, CD14, CD16, CD20, CD23, CD34, CD123), NKp44+CD127+CD218+ lymphocytes. Positive/negative gating based on clearly grouped populations, historical-determined expression, and the use of internal controls (Supplementary Fig. 8). A threshold of 100 collected events in the parent population was utilized for all subset expression analysis.
Ortiz A.M., Flynn J.K., DiNapoli S.R., Vujkovic-Cvijin I., Starke C.E., Lai S.H., Long M.E., Sortino O., Vinton C.L., Mudd J.C., Johnston L., Busman-Sahay K., Belkaid Y., Estes J.D, & Brenchley J.M. (2018). Experimental Microbial Dysbiosis Does Not Promote Disease Progression in SIV-Infected Macaques. Nature medicine, 24(9), 1313-1316.
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4

Activation of CD134 in PBMCs

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Stimulated PBMCs were further co-cultured with an isotype control (cetuximab, Southampton General Hospital pharmacy), anti-CD134 (SAP25–29 h1, in-house), deglycosylated anti-CD134 (in-house), anti-CD134 F(ab′)2 (in-house, produced as previously described37 (link)), multimeric CD134L (Caltag) or control ligand (Caltag) for 6 hours. For IFNγ (B27, BD Bioscience) and TNFα (Mab11, eBioscience) intracellular staining, human cells were fixed and permeabilised as per manufacturer’s protocol (BD Bioscience) after 6 hours co-culture with brefeldin A (BD Bioscience). For assessment of NK degranulation, PBMCs were incubated with CD107a-PE (H4A3, BD Bioscience) and monensin (BD Bioscience) for 6 hours.
Turaj A.H., Cox K.L., Penfold C.A., French R.R., Mockridge C.I., Willoughby J.E., Tutt A.L., Griffiths J., Johnson P.W., Glennie M.J., Levy R., Cragg M.S, & Lim S.H. (2018). Augmentation of CD134 (OX40)-dependent NK anti-tumour activity is dependent on antibody cross-linking. Scientific Reports, 8, 2278.
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