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Figure 1 | BMC Cancer

Figure 1

From: Novel anti-HER2 monoclonal antibodies: synergy and antagonism with tumor necrosis factor-α

Figure 1

Strong binding of anti-HER2 antibodies to ERBB2 -amplified breast cancer cell lines. ERRB2 amplification was analyzed by FISH analysis. Cells were harvested at confluence, fixed and subjected to dual-color FISH using Texas Red-labeled ERBB2 (red) and FITC-labeled chromosome 17 α-satellite CEP17 (green) DNA probes (left panel). For generation of anti-HER2 monoclonal antibodies, mice were immunized with SK-BR-3 cells, followed by a recombinant protein composed of extracellular domain of HER2 fused to human IgG1 Fc domain (HER2 ECD). Antibodies were purified from conditioned hybridoma cell culture media by affinity chromatography. For immunofluorescence assay, cells were seeded on coverslips in 6-well plates, incubated 36 h in cell culture medium, fixed, and permeabilized. Fixed cells were incubated with anti-HER2 antibodies, and the primary antibody binding was detected using Alexa 488- conjugated anti-mouse IgG (BH1, BH2, BH5, BH6, BH7 panels; green). Nuclei were counterstained with DAPI (blue) and pictures were acquired using Zeiss Axio Imager.A1 microscope.

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