Surface levels of HLA‐I molecules were determined on glioblastoma cell lines using the following antibodies: anti‐HLA‐(A,B,C)‐PE (clone W6/32), anti‐HLA‐E‐PE (clone 3D12), anti‐HLA‐F‐PE (clone 3D11/HLA‐F), and anti‐HLA‐G‐PE (clone 87G) (all from Biolegend). PE mouse IgG1 (clone MOPC‐21) or PE mouse IgG2a (clone MOPC‐173) antibodies (both from Biolegend) were employed as isotype controls. 7‐AAD (Immunostep) was used to discriminate viable cells. Cells were analyzed in a Cytoflex S flow cytometer with CytExpert 2.3 software (Beckman Coulter). For the analysis of surface expression, raw MFI values were corrected by subtracting isotype values and, where indicated, normalized to the control condition as a fold induction.
Anti hla g pe clone 87g
Manufactured by BioLegend
Anti‐HLA‐G‐PE (clone 87G) is a monoclonal antibody labeled with phycoerythrin (PE) that binds to HLA‐G, a non‐classical major histocompatibility complex (MHC) class I molecule. This antibody can be used for the detection and analysis of HLA‐G expression on cells by flow cytometry.
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2 protocols using anti hla g pe clone 87g
1
Quantifying HLA-I Molecule Expression
2
Profiling ILT2 and Ligands in Lymphocytes
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For analyzing the expression of ILT2 and its ligands in different lymphocyte subsets, 1 × 106 PBMCs obtained from CLL patients and healthy donors were incubated with the corresponding antibodies at room temperature for 20 min. Anti-CD3-FITC, anti-CD56-APC, and anti-CD19-APC antibodies (all from Immunostep), anti-CD3-PECy7 and anti-CD56-PECy7 antibodies (eBioscience), and appropriate isotype-matched controls were used to define the immune subsets. Anti-ILT2-PE (clone HP-F1, eBioscience), anti-HLA-F rabbit antibody (Abgent), anti-HLA-G-PE (clone 87G Biolegend), anti-HLA-E-PE (clone 3D12 Biolegend), and a pan-anti-HLA-I antibody (clone W6/32) were used to measure the expression of ILT2 and its ligands in NK cells and B cells. NK cells were defined as CD3−CD56+, and B cells as CD19+. Cells were analyzed on a BD Biosciences FACS Canto II cytometer (Beckton Dickinson).
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