Whole blood samples were incubated with HIV-1 gag, pol and env peptide pools respectively at 37 °C for 5 h. Brefeldin-A (10 μg/ml, Sigma), GolgiStop (5 μg/ml, BD Biosciences) and anti-CD107a APC (clone H4A3, 20 μl/ml, BD Biosciences) were added immediately to the cell medium and incubated for 6 h. Samples incubated with phorbol-12-myristate-13-acetate (PMA) (1 mg/ml) plus ionomycin (1 mg/ml) were used as positive controls and samples incubated with DMSO (1 mg/ml, Sigma) were used as negative controls. Samples were surface-stained with live/dead dye Amcyan (Invitrogen, Carlsbad, CA, USA), anti-CD3-PerCP (clone SK7) and anti-CD56-PE (clone B159), and then lysed, permeabilized and intracellularly stained with anti-IFNγ Alexa Fluor700 (clone B27, BD Biosciences) and anti-CD107a APC (clone H4A3). All data were acquired on BD FACS Fortessa (BD Biosciences, San Jose, CA, USA) and analyzed by FlowJo software (Treestar, Ashland, OR, USA).
Anti cd107a apc clone h4a3
Manufactured by BD
Sourced in United States
Anti-CD107a-APC (clone H4A3) is a monoclonal antibody that binds to the CD107a (LAMP-1) surface antigen. The antibody is conjugated with the APC (Allophycocyanin) fluorescent dye.
Automatically generated - may contain errors
Lab products found in correlation
Brefeldin a, by Merck Group (2 mentions)
Ionomycin, by BD (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Dnase, by Roche (1 mentions)
Il 12, by R&D Systems (1 mentions)
Il 18, by R&D Systems (1 mentions)
Monensin, by BD (1 mentions)
Il 12, by Immunotools (1 mentions)
Granzyme b, by Mabtech (1 mentions)
Goat anti human ig pe, by Southern Biotech (1 mentions)
Anti egfr mab 425, by Merck Group (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
Annexin 5 fitc, by Immunotools (1 mentions)
Interleukin 6 (il 6), by Immunotools (1 mentions)
Interleukin 4 (il 4), by Immunotools (1 mentions)
Il 1β, by Immunotools (1 mentions)
Gm csf, by Immunotools (1 mentions)
Recombinant human ifn γ, by Immunotools (1 mentions)
Amcyan, by Thermo Fisher Scientific (1 mentions)
Anti cd3 percp clone sk7, by BD (1 mentions)
3 protocols using anti cd107a apc clone h4a3
1
HIV-1 Antigen-Specific T-Cell Response Assay
2
Activation of Frozen Immune Cells
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Frozen perfusate cells were thawed, washed and incubated in RPMI + 10% FCS in the presence of 10 U/ml DNAse (Roche) for 45 min at 37 °C. After washing, cells were cultured with either 5 ng/ml IL-12 and 50 ng/ml IL-18 (both R&D), 3 ng/ml PMA and 100 ng/ml Ionomycin (both Sigma, low PMA+Ionomycin) or 25 ng/ml PMA and 1 μg/ml Ionomycin for 4 h at 37 C. Anti-CD107a-APC (clone H4A3, BD Bioscience) was present during culture. Brefeldin A (Sigma) and Monensin (BD Bioscience) were added for the last 3 h of stimulation.
3
Detailed Immunophenotyping of T-Cell Responses
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Goat anti-human Ig-PE (Southern Biotech), anti-PD-L1-APC (clone 29E.2A3, BioLegend), anti-EGFR-FITC (clone 528, Santa Cruz Biotechnology), anti-CD107a-APC (clone H4A3, BD Pharmingen), anti-CD137-PE (clone 4B4), anti-IFN-γ-PerCP-Cyanine5.5 (clone 4S.B3,), anti-CD3-PerCP-Cyanine5.5 (clone OKT-3, all from eBioscience), and anti-CD3-FITC (clone Ucht1), anti-CD8-FITC, APC (clone HIT8a), anti-CD56-PE (clone B-A19), anti-CD14-FITC, PE (clone MEM-15), anti-CD25-FITC, APC (clone MEM-181), anti-HLA-DR-FITC, PE (clone MEM-12), mouse IgG1-FITC, PE, Mouse IgG2b-APC, Annexin-V-FITC (all from Immunotools). Recombinant human IFN-γ, TNF-α, PGE2, GM-CSF, IL-1β, IL-4, IL-6, IL12 and anti-CD3 mAb UCHT-1 were from Immunotools. PD-L1-blocking mAb was from BPS Bioscience. Anti-EGFR mAb 425 was from Merck. Cetuximab was obtained from the Department of Hospital Pharmacy, UMCG, The Netherlands. Secretion of cytokines by T cells was measured using appropriate ELISA kits (IFN-γ from eBioscience and granzyme B from Mabtech).
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