Measurement of the surface HER2 expression was performed with HER2-overexpressing human breast cancer cell line (SKBR3). 400 ng/mL of SB3 and reference product were incubated with SKBR3 at 37°C, 5% CO2 for 24 h. Following incubation, cells were fixed by formalin and labeled with PE-conjugated IgG (R&D Systems) or PE anti-human CD340 (ErbB2/HER2) antibody (BioLegend). The surface HER2 level was measured by flow cytometry analysis (FACSVerse™, BD Biosciences, San Jose, CA, USA). This quantified the PE fluorescence, the subsequent reduction of which showed SB3- or reference product-induced reduction of surface HER2.
Pe anti human cd340 erbb2 her2 antibody
Manufactured by BioLegend
Sourced in United States
The PE anti-human CD340 (ErbB2/HER2) antibody is a laboratory reagent used for the detection and analysis of the CD340 (ErbB2/HER2) receptor. It is conjugated with the fluorescent dye Phycoerythrin (PE), which allows for the identification and quantification of CD340-positive cells through flow cytometry or other fluorescence-based techniques.
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Lab products found in correlation
Facsverse, by BD (1 mentions)
Docetaxel taxotere, by Sanofi (1 mentions)
Cytotox glo, by Promega (1 mentions)
Recombinant human heregulin β1, by Thermo Fisher Scientific (1 mentions)
Celltrace carboxyfluorescein succinimidyl ester cfse cell proliferation kit, by Thermo Fisher Scientific (1 mentions)
Celltiter blue, by Promega (1 mentions)
Human vegf quantikine elisa kit, by R&D Systems (1 mentions)
Caspase glo 3 7 assay kit, by Promega (1 mentions)
Aldefluor assay kit, by STEMCELL (1 mentions)
3 protocols using pe anti human cd340 erbb2 her2 antibody
1
Quantifying HER2 Expression in SKBR3 Cells
2
Evaluation of ErbB2/HER2 Biosimilar Candidates
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The reference products in this study were selected randomly from 154 lots of EU- and US-sourced reference product. The CellTiter-Blue® and CytoTox-Glo® kits were obtained from Promega (Madison, WI, USA). The phycoerythrin (PE) anti-human CD340 (ErbB2/HER2) antibody was obtained from Biolegend (San Diego, CA, USA). The ErbB2/HER2 enzyme-linked immunosorbent assay (ELISA) kit was obtained from R&D Systems (Minneapolis, MN, USA), the Phospho-Akt (Ser473) ELISA kit was obtained from Cell Signaling Technology (Danvers, MA, USA), and the Human VEGF Quantikine® ELISA kit was obtained from R&D Systems. The CellTrace™ carboxyfluorescein succinimidyl ester (CFSE) Cell Proliferation Kit was obtained from Thermo Fisher Scientific (Waltham, MA, USA), the docetaxel (Taxotere®) was obtained from Sanofi-Aventis (Paris, France), and the PathHunters® Flash Detection Kit was obtained from DiscoverX (Fremont, CA, USA). The recombinant human heregulin-β1 was obtained from Peprotech (Seoul, South Korea) and the Caspase-glo 3/7 assay kit was obtained from Promega.
3
ALDH Activity Assay in Cancer Cells
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ALDH activity was measured using an ALDEFLUOR assay kit (StemCell Technologies, Vancouver, Canada) in OE19, N87, OE33, and GTR0455 cells according to the manufacturer’s instructions. Briefly, 7.5 × 105 cells were suspended in ALDEFLUOR assay buffer containing the ALDH substrate BODIPY-aminoacetaldehyde (BAAA) and incubated for 1 h at 37 °C. A specific inhibitor of ALDH, diethylaminobenzaldehyde (DEAB), was used to distinguish between ALDH-positive (ALDH +) and ALDH-negative (ALDH-) cell subsets. For double-marker immunofluorescence staining, cells were incubated for 1 h at 37 °C both with an ALDEFLUOR assay kit (StemCell Technologies) and with PE-anti-human CD340 (erbB2/HER-2) antibody (1:100) (BioLegend, San Diego, CA, USA) to analyze HER2 expression. The cells were analyzed using a BD LSR II FACScanto cytometer (BD Biosciences, Franklin Lakes, NJ, USA). In all experiments, the data were processed using the FlowJo software package.
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