377 hb cf  (R&D Systems)

Journal: Life sciences

Article Title: High ErbB3 activating activity in human blood is not due to circulating neuregulin-1 beta

doi: 10.1016/j.lfs.2020.117634

Figure Lengend Snippet: A. Graphical representation of ErbB1-4 receptors expression in a human microvascular endothelial cell line (HMEC1, positive control), a human monocytic cell line (THP1, negative control) and a human breast cancer cell line (MCF-7). B. Flow cytometric analysis of cell surface expression of ErbB receptors in MCF-7 cells. The open histogram depicts the specific ErbB antibody and the gray histogram represents the isotype-matched control. The Y-axis shows cell count; the X-axis indicates fluorescence intensity. C. Effect of 10 ng/ml neuregulin-1(NRG-1) on phosphorylation of ErbB3 in MCF-7 cells. The horizontal lines on the left are the molecular weight ladder. The ErbB3 molecular weight is 180kD. D. Graphical representation of data from western blot analysis of NRG-1 induced phosphorylation of ErbB3. Data are shown as mean±SEM; n=3.

Article Snippet: Reagents An extracellular domain of recombinant human neuregulin-1 (377-HB/CF), recombinant human epidermal growth factor (EGF, 236-EG), recombinant human betacellulin (BTC, 261-CE), and recombinant human heparin-binding epidermal growth factor (HB-EGF, 259-HE) were purchased from Bio-techne/R&D Systems and reconstituted in phosphate-buffered saline.

Techniques: Expressing, Positive Control, Negative Control, Cell Counting, Fluorescence, Molecular Weight, Western Blot

Journal: Breast Cancer Research : BCR

Article Title: Polyfunctional anti-human epidermal growth factor receptor 3 (anti-HER3) antibodies induced by HER3 vaccines have multiple mechanisms of antitumor activity against therapy resistant and triple negative breast cancers

doi: 10.1186/s13058-018-1023-x

Figure Lengend Snippet: Human epidermal growth factor receptor 3 ( HER3)-specific antibody responses are induced by adenovirus encoding full length human HER3 (Ad-HER3) in vivo. a Binding of vaccine-induced antibody (VIA) in serum from mice immunized with Ad-HER3 (HER3-VIA), Ad-lacZ (lacZ-VIA), and Ad-green fluorescence protein (GFP-VIA). Serum at dilutions presented were mixed with the HER3-4 T1 cell line or wild-type 4 T1 and binding of antibody was identified with near infrared (nIR) dye-conjugated anti-mouse IgG and detected by a LI-COR Odyssey Imager. The difference in fluorescence intensity between HER3-4 T1 and 4 T1 is graphed. b Binding of HER3-VIA to breast cancer cell lines: a panel of human breast cancer cell lines were incubated with dilutions of the HER3-VIA, washed, and then mixed with a phycoerythrin (PE)-conjugated secondary antibody. HER3-VIA binding was analyzed based on fluorescence activated cell sorting analysis and mean fluorescence intensity was reported. c Flow cytometric analysis was used to identify percentage of BT474 cells able to bind HER3-VIA (1:100 dilution). d Binding of His-tagged heregulin to HER3 receptor was measured by pre-incubating BT474 cells with heregulin (0, 10, 100 nM), or HER3-VIA (1:100) for 10 min on ice, followed by incubation with His-tagged heregulin (100 nM) for 10 min. Receptor-bound His-tagged heregulin was visualized by staining with PE-conjugated anti-His Tag antibody. Mean fluorescence intensity is shown in each histogram

Article Snippet: Heregulin (377-HB/CF), heregulin with a C-terminal 6-His tag (5898-NR) and allophycocyanin (APC)-conjugated anti-His Tag antibody (IC050A) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: In Vivo, Binding Assay, Fluorescence, Incubation, FACS, Staining