For murine T cell work, we used the following BD Bioscience antibodies: mouse Fc-Block, anti-CD4 (clone RM4-5), anti-CD8a (clone 53-6.7), anti-IFNγ (clone XMG1.2), and anti-TNFα (clone MP6-XT22). For human T-cell staining, we used the following BD Bioscience antibodies: anti-CD4 (clone RPA-T4), anti-NGFR (clone C40-1457), anti-IFNγ (clone B27), and anti-TNFα (clone MAb11). The anti-CD8a clone OKT8 from eBiosciences was also used. Recombinant human HER2-Fc (R&D Systems) was used to stain for HER2-CAR expression on both murine and human T cells and detected with goat-anti-human IgG (Jackson ImmunoResearch, West Grove, PA). Intracellular staining were performed using the cytofix/cytoperm reagent and associated protocol (BD Biosciences). T-cell viability was evaluated using the Molecular Probes LIVE/DEAD Fixable Near-IR kit (Life Technologies). Results were acquired on a FACSCanto or LSRII (BD Biosciences) and analyzed using FlowJo software (FlowJo, Ashland, OR).
Anti tnfα clone mp6 xt22
Manufactured by BD
Sourced in United States
The Anti-TNFα (clone MP6-XT22) is a laboratory reagent used for the detection and quantification of tumor necrosis factor-alpha (TNF-α) in various biological samples. It is a monoclonal antibody that specifically binds to TNF-α, a pro-inflammatory cytokine involved in immune responses and inflammatory processes.
Automatically generated - may contain errors
Lab products found in correlation
Facscanto, by BD (1 mentions)
Cytofix cytoperm reagent, by BD (1 mentions)
Mouse fc block, by BD (1 mentions)
Anti ifnγ clone b27, by BD (1 mentions)
Anti tnfα clone mab11, by BD (1 mentions)
Live dead fixable dead cell stain kit, by Thermo Fisher Scientific (1 mentions)
Anti cd3, by BD (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
Anti cd8, by BD (1 mentions)
Brefeldin a, by BD (1 mentions)
Cytofix cytoperm kit, by BD (1 mentions)
Anti il 2 jes6 5h4, by BD (1 mentions)
Anti cd4 clone rm4 5, by BD (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Monensin, by Merck Group (1 mentions)
Monensin, by BD (1 mentions)
Cytofix cytoperm fixation and permeabilization kit, by BD (1 mentions)
Phorbol 12 myristate, by Merck Group (1 mentions)
Anti ifn γ clone xmg1, by BD (1 mentions)
3 protocols using anti tnfα clone mp6 xt22
1
Murine and Human T Cell Immunophenotyping
2
Cytokine Profiling of Mouse Splenocytes Exposed to SARS-CoV-2 Peptides
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The mouse splenocytes were stimulated at 37 °C for 20 h with or without the PepMixTM SARS-CoV-2(S-RBD) peptide pool (JPT Peptide Technology, Berlin, Germany) (2 µg/mL) in the presence of 10 μg/mL brefeldin A (BD Biosciences, San Jose, CA, USA). Cells were stained with a yellow LIVEDEAD fixable dead cell stain kit (Invitrogen, Waltham, MA, USA) and anti-CD3 (Clone 500A2, BD), anti-CD4 (Clone RM4-5, BD), and anti-CD8 (Clone 53-6.7, BD) antibodies. The cells were subsequently fixed and permeabilized using the Cytofix/Cytoperm kit (BD Biosciences, San Jose, CA, USA) and stained with anti-TNF-α (clone MP6-XT22, BD, 1:100) and anti-IL-2 (JES6-5H4, BD, 1:100) antibodies. The cells were acquired on a CytoFLEX (Beckman Coulter, Brea, CA, USA) and analyzed with Flowjo software (Tree Star, Ashland, OR, USA).
3
Cytokine Analysis in Murine Spleen Cells
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Spleen cells from different groups of BALB/c mice were analyzed for cytokine content as previously described [48 (link)]. Briefly, T cells were seeded at 106/mL in RPMI 1640–10% FBS, and stimulated with 10 ng/mL phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich) and 500 ng/mL ionomycin (Sigma-Aldrich) in the presence of monensin (BD) during 4-h incubation at 37 °C, and then treated with Cytofix/Cytoperm fixation and permeabilization kit (BD). For macrophage stimulation, spleen cells were seeded at 106/mL in RPMI 1640–10% FBS, and stimulated with LPS 100 ng/mL (Sigma-Aldrich) in the presence of monensin during 4 h incubation at 37 °C. The permeabilized cells were intracellularly stained with the PE-conjugated cytokine-specific mAbs anti-IFNγ (clone XMG1.2), and anti-TNFα (clone MP6-XT22), all from BD; or isotype-matched irrelevant mAbs from BioLegend.
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