reddot 2 dyes  (Reddot Biotech)

Journal: PLOS One

Article Title: Inhibition of HER2 signaling and breast cancer cell growth with a novel antibody targeting HER2 ECD III/IV

doi: 10.1371/journal.pone.0338127

Figure Lengend Snippet: A. Competitive ELISA assessing the binding of His-tagged IgGs to immobilized HER2 in the presence or absence of saturating concentrations of non-His-tagged IgGs, detected with an anti-His-HRP antibody. B. Saturation binding curves of trastuzumab and pertuzumab to HER2 on live MCF7 cells. Cells were incubated with antibodies (0-400 nM) for 30 min and detected with an AF647-conjugated goat anti-human secondary antibody via flow cytometry. C. Competitive binding of of AF647-labeled trastuzumab or pertuzumab (0-300 nM) to MCF7 cells pre-saturated with 267 nM of unlabeled trastuzumab and pertuzumab, analyzed by flow cytometry. D. Binding of AF647-conjugated m66 or m75 to MCF7 cells after pre-incubation with 267 nM trastuzumab or pertuzumab. E. Binding of AF647-conjugated rabbit antibodies to MCF7 cells pre-saturated with 267 nM of trastuzumab and pertuzumab, evaluated by flow cytometry. T, trastuzumab; P, pertuzumab. Error bars, SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001, T-test.

Article Snippet: Briefly, each antibody was reacted with 20 molar equivalents of AF647 NHS ester (Biotium, 30–3007) for 2 h at room temperature protected from light.

Techniques: Competitive ELISA, Binding Assay, Incubation, Flow Cytometry, Labeling