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Annexin 5 stain

Manufactured by BD
Sourced in United States

Annexin V is a calcium-dependent phospholipid-binding protein that has a high affinity for phosphatidylserine (PS). Annexin V stain is used to detect and quantify apoptotic cells in flow cytometry and fluorescence microscopy applications.

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2 protocols using annexin 5 stain

1

Evaluation of Cisplatin, Cetuximab, and Panitumumab Efficacy

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Cisplatin was manufactured by Fresenius Kabi (Lake Zurich, IL) and obtained from the pharmacy at the University of Pittsburgh Cancer Institute. Cetuximab was from Brystol-Myers Squibb (New York, NY), and panitumumab was from Amgen (Thousand Oaks, CA). Water-soluble gefitinib and SB203580 were obtained from Tocris Bioscience. Anti-human antibodies used for flow cytometry staining included Alexa 488-p-S727-STAT1er727, PE-pSTAT1Tyr701, Alexa 647-STAT1 N terminus, Pacific Blue-p-p38 and corresponding isotype controls, all from BD Biosciences. Anisomycin, antibodies to p-S727-STAT1 and p-Y701-STAT1 for immunohistochemistry of tumor specimens, and antibodies for Western blots (total STAT1 primary and anti-rabbit secondary) were obtained from Cell Signaling Technology. Annexin V stain and Annexin buffer were obtained from BD Biosciences. Zombie Aqua viability dye was obtained from BioLegend and used at a dilution of 1/500. Lipopolysaccharide and β-actin antibody were obtained from Sigma. The APO-BrdU TUNEL assay kit for flow cytometry was from Life Technologies.
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2

Apoptosis Assessment via Annexin V

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Apoptotic activity due to neuronal induction was assessed using Annexin V stain (Catalog # 556547, BD Pharmingen, USA). Neurospheres from each group were gently dissociated into single-cell format, resuspended in 1X binding buffer, and then incubated with FITC-conjugated Annexin V and propidium iodide for 15 minutes, according to the manufacturer’s recommended protocol. Apoptotic activity was observed under a confocal microscope, and quantitative results were obtained by flow cytometry analysis.
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