Human ErbB2/Her2 (Research Grade Trastuzumab Biosimilar) Antibody

Detection of ErbB2/Her2 in MCF-7 human cell line by Flow Cytometry. MCF-7 human cell line was stained with Human Anti-Human ErbB2/Her2 (Research Grade Trastuzumab Biosimilar) Monoclonal Antibody (Catalog # MAB9589, filled histogram) or Human IgG Control Antibody (1-001-A, open histogram) followed by APC-conjugated Anti-Human IgG Secondary Antibody (Catalog # F0135). View our protocol for Staining Membrane-associated Proteins.

Detection of ErbB2/Her2 in MDA-MB-453 human cell line by Flow Cytometry. MDA-MB-453 human cell line was stained with Human Anti-Human ErbB2/Her2 (Research Grade Trastuzumab Biosimilar) Monoclonal Antibody (Catalog # MAB9589, filled histogram) or Human IgG Control Antibody (1-001-A, open histogram) followed by APC-conjugated Anti-Human IgG Secondary Antibody (Catalog # F0135). View our protocol for Staining Membrane-associated Proteins.

Inhibition of Cell Proliferation by ErbB2/Her2 (Research Grade Trastuzumab Biosimilar) Antibody. Human Anti-Human ErbB2/Her2 (Research Grade Trastuzumab Biosimilar) Monoclonal Antibody (Catalog # MAB9589) inhibits proliferation in the SK-BR-3 human breast cancer cell line as measured by Resazurin (Catalog # AR002). The ED₅₀ for this effect is typically 15-75 ng/mL.

NKs cells are potent killers in ADCC assays with SKOV-3 or Raji cells (A, B) ADCC assays with SKOV-3 and Raji targets. NK cells were cultured as described in the adjacent figure for 13 days. Prior to setting up the ADCC assay, CD56+CD16+ levels were ascertained to be >30% on NKs using Human Fc gamma RIII (CD16) PerCP-conjugated Antibody (FAB2546C); in our hands, this is the minimum level required for killing via ADCC. Target cells were labeled with Mito Mark Green (SKOV-3 or Raji), and Effectors/NKs were labeled with Janelia Fluor 646 as described (Figure 1). (A) SKOV-3 cells were preincubated with 5 ug/mL Anti-Her2/Trastuzumab (Catalog # MAB9589); (B) Raji cells were preincubated with 10 ug/mL Anti-CD20/Rituximab (MAB9630), both for 30 minutes at room temperature prior to setting up the ADCC assay with the NKs. 0.5:1 E:T ratios were used for the SKOV-3/Trastuzumab model, and NKs and SKOV-3 cells were co-incubated for 2 hours at 37°C. 2:1 E:T ratios were used for the Raji/Rituximab model, and NKs and Raji cells were co-incubated for 2.5 hours at 37°C. In both models, the cells were harvested, stained with antibodies for Human CD3 Alexa Fluor 405 and Human CD56 Alexa Fluor 700, and analyzed by flow cytometry. Importantly, non-NKs (JF 646-CD56- cells) are shown in the Mito Mark dye graphs that depict killing. These results are representative of 3+ experiments.