Immature DCs were generated by culturing healthy monocytes in an X-vivo 15 free serum medium (Lonza, Walkersville, MD) supplemented with 1000 U/ml GM-CSF (Miltenyi) and 500 U/ml IL-4 (Miltenyi) for 6 days. The maturation of DCs was induced by adding to the medium one of the following stimulants: 100 ng/ml LPS (Sigma-Aldrich, St-Louis, MO), 0.5 μg/ml mouse IgG2a anti-human monoclonal HLA class I antibody clone W6/32 (Sigma-Aldrich), 0.5 μg/ml mouse IgG2a anti-human monoclonal HLA-DR, DP, DQ clone Tü39 (BD Biosciences, San Diego, CA), or 0.5 μg/ml mouse IgG2a isotype control (Sigma-Aldrich and BD Biosciences). The supernatants were collected at the end of the DC maturation stage for all samples.
Mouse igg2a anti human monoclonal hla class 1 antibody clone w6 32
Manufactured by Merck Group
The Mouse IgG2a anti-human monoclonal HLA class I antibody clone W6/32 is a laboratory reagent used for the detection and study of human major histocompatibility complex (MHC) class I molecules. It recognizes a monomorphic epitope on the HLA-A, -B, and -C heavy chains, allowing for the identification and analysis of HLA class I expression on cells.
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Lab products found in correlation
Mouse igg2a isotype control, by Merck Group (1 mentions)
X vivo 15 serum free medium, by Lonza (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Gm csf, by Miltenyi Biotec (1 mentions)
Interleukin 4 (il 4), by Miltenyi Biotec (1 mentions)
Thrombin, by Thermo Fisher Scientific (1 mentions)
Tacrolimus, by Enzo Life Sciences (1 mentions)
2 protocols using mouse igg2a anti human monoclonal hla class 1 antibody clone w6 32
1
Dendritic Cell Maturation Protocols
2
Monoclonal Antibody-Induced Endothelial Stimulation
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Mouse IgG2a anti-human monoclonal HLA class I antibody clone W6/32 (Sigma-Aldrich, St Louis, MO), mouse IgG2a anti-human monoclonal HLA-DR, DP, DQ clone Tü39 (BD Biosciences, San Diego, CA), and clone WR18 (ThermoFisher Scientific, Rockford, IL) at 1 or 2 μg/mL were used to stimulate endothelial cells for 10 minutes or 1 hour. The concentrations of anti-HLA antibodies used are the expected physiological concentrations in human blood, as measured after the purification of DSAs from patients using specific immunoaffinity columns. 17 Stimulation with histamine (Prepotech, Rocky Hill, NJ) at 1.0 mmol/L for 10 minutes or thrombin at 1 U/mL (ThermoFisher Scientific) for 1 hour was used as a positive control. Tacrolimus was obtained from Enzo Life Sciences Inc. (Farmingdale, NY). Assessment of cell viability and the methods used to test cyclosporine, prednisone, and mycophenolic acid are described as Supplemental Material.
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