mouse igg2b (clone mpc-11) antibody  (Bio X Cell)

Journal: Cancer Cell

Article Title: TIM-3 blockade in diffuse intrinsic pontine glioma models promotes tumor regression and antitumor immune memory

doi: 10.1016/j.ccell.2023.09.001

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Article Snippet: InVivoMAb anti-IgG2b (MPC-11) , BioXCell , Cat#BE0086; RRID: AB_1107791.

Techniques: Virus, shRNA, Recombinant, BrdU Cell Proliferation Assay, Enzyme-linked Immunospot, Expressing, Control, Mouse Assay, Software

Journal: PLOS ONE

Article Title: THP-1 cells transduced with CD16A utilize Fcγ receptor I and III in the phagocytosis of IgG-sensitized human erythrocytes and platelets

doi: 10.1371/journal.pone.0278365

Figure Lengend Snippet: THP-1 cells were differentiated to macrophages by treatment with PMA (100 ng/mL) as described in the methods. (A) THP-1 cells. (B) THP1-CD16A cells. Opsonization: (-) indicates erythrocytes were non-opsonized (incubated with phosphate buffered saline), (+) indicates erythrocytes were opsonized with a polyclonal anti-human RhD antibody (WinRho SDF TM ). The contribution of each FcγR to the phagocytosis was evaluated using Fc region deglycosylated blocking antibodies (final concentration of 10 μg/mL each): anti-FcγRI (clone 10.1), anti-FcγRIIA (clone IV.3), anti-FcγRIIA/B/C (clone AT10), or anti-FcγRIIIA (clone 3G8). The deglycosylated mouse IgG1 (clone MOPC-21) and deglycosylated mouse IgG2b (clone MPC-11) were used in combination as isotype controls (final concentration of 10 μg/mL each). The phagocytic index was calculated as the number of erythrocytes engulfed per 100 macrophages. Data are presented as the mean ± the standard deviation of five independent experiments. For panels A and B, the statistical analysis was performed with a one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons test (****: p <0.001).

Article Snippet: Purified mouse anti-human CD32 (clone IV.3), FITC-conjugated mouse anti-human CD32 (clone IV.3), and FITC-conjugated mouse IgG2b isotype control (clone MPC-11) were purchased from Stemcell Technologies Inc. (British Columbia, Canada).

Techniques: Incubation, Blocking Assay, Concentration Assay, Standard Deviation

Journal: PLOS ONE

Article Title: THP-1 cells transduced with CD16A utilize Fcγ receptor I and III in the phagocytosis of IgG-sensitized human erythrocytes and platelets

doi: 10.1371/journal.pone.0278365

Figure Lengend Snippet: Three different images are shown of IgG-opsonized platelets incubated with THP-1-CD16A macrophages. Platelets were labelled with 5- chloromethylfluorescein diacetate (CMFDA) (green) before phagocytosis. Non-phagocytosed platelets were differentiated after phagocytosis using an AF647-conjugated anti-human CD42a antibody (red). Platelets were additionally defined by size (roughly 1.5 to 3.5 μm) to distinguish them from internalized microparticles or platelet aggregates. THP-1-CD16A macrophages were observed by spinning-disc confocal microscopy under 63x objective oil immersion with differential interference contrast and DAPI stain on a Quorum multi-modal imaging system (Quorum Technologies, Ontario, Canada). Four images were taken at the center of each well with Z-stacking. Phagocytosis was quantified using Imaris v9.6.0. Scale bar = 3 μm. White arrows indicate examples of phagocytosed platelets.

Article Snippet: Purified mouse anti-human CD32 (clone IV.3), FITC-conjugated mouse anti-human CD32 (clone IV.3), and FITC-conjugated mouse IgG2b isotype control (clone MPC-11) were purchased from Stemcell Technologies Inc. (British Columbia, Canada).

Techniques: Incubation, Confocal Microscopy, Staining, Imaging

Journal: PLOS ONE

Article Title: THP-1 cells transduced with CD16A utilize Fcγ receptor I and III in the phagocytosis of IgG-sensitized human erythrocytes and platelets

doi: 10.1371/journal.pone.0278365

Figure Lengend Snippet: THP-1-CD16A cells were differentiated to macrophages by treatment with PMA (100 ng/mL) as described in the Materials and methods. (A) Magnitude of macrophage phagocytosis of IgG-opsonized platelets. Opsonization: (-) indicates platelets were non-opsonized (incubated with normal human serum), (+) indicates platelets were opsonized with the monoclonal antibody A2A9 (5 μg/mL). The phagocytic index was calculated as the number of platelets engulfed per 100 macrophages. Data are presented as the mean ± the standard deviation of eight independent experiments. The statistical analysis was performed by the non-parametric Mann-Whitney T-test (****: p <0.0001). (B) The contribution of each FcγR to the phagocytosis was evaluated using Fc region deglycosylated blocking antibodies (final concentration of 10 μg/mL each): anti-FcγRI (clone 10.1), anti-FcγRIIA/B/C (clone AT10), or anti-FcγRIIIA (clone 3G8). The deglycosylated mouse IgG1 (clone MOPC-21) was used as isotype control (final concentration of 10 μg/mL). Percent phagocytosis was assessed relative to macrophages without any antibody treatment of the THP-1-CD16A cells. Data are presented as the mean ± the standard deviation of five independent experiments. The statistical analysis was performed with a one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons test (***: p <0.001).

Article Snippet: Purified mouse anti-human CD32 (clone IV.3), FITC-conjugated mouse anti-human CD32 (clone IV.3), and FITC-conjugated mouse IgG2b isotype control (clone MPC-11) were purchased from Stemcell Technologies Inc. (British Columbia, Canada).

Techniques: Incubation, Standard Deviation, MANN-WHITNEY, Blocking Assay, Concentration Assay