Annexin 5 alexafluor 647 conjugated antibody

Manufactured by Thermo Fisher Scientific

The Annexin V AlexaFluor 647-conjugated antibody is a fluorescently labeled protein that binds to phosphatidylserine. It is commonly used in flow cytometry and fluorescence microscopy applications to detect and quantify apoptotic cells.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Live dead fixable violet dead cell dye, by Thermo Fisher Scientific (1 mentions) Annexin 5 binding buffer, by BD (1 mentions)

Close spelling variants of this product

AlexaFluor-647 antibody Annexin V antibody AlexaFluor antibody Annexin V

2 protocols using annexin 5 alexafluor 647 conjugated antibody

Comprehensive Cell Surface and Apoptosis Analysis

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All flow cytometry analysis was performed on a LSR Fortessa analyser (Becton Dickinson, Franklin Lakes, NJ). For analysis of BAP1 expression cells were stained with primary antibody to BAP1 (Santa Cruz Biotechnology Cat# sc-28383, RRID:AB_626723; 1:50) and then with an AlexaFluor 488-conjugated anti-mouse antibody (Thermo Fisher Scientific Cat# A-21202, RRID:AB_141607; 1:200). For analysis of apoptosis and cell death all floating and adherent cells were harvested and stained with an Annexin V AlexaFluor 647-conjugated antibody (Thermo Fisher Scientific Cat# A23204, RRID:AB_2341149) and 4’, 6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich, 200 μg/ml). For analysis of DR4 and DR5 expression on cell surface cells were stained with PE-conjugated antibody (DR4 - BioLegend, San Diego, CA) Cat# 307205, RRID:AB_314669, DR5 - BioLegend Cat# 307405, RRID:AB_314677, Isotype control - Biolegend #400112; 1:100). FlowJo software was used to analyse all data.

Kolluri K.K., Alifrangis C., Kumar N., Ishii Y., Price S., Michaut M., Williams S., Barthorpe S., Lightfoot H., Busacca S., Sharkey A., Yuan Z., Sage E.K., Vallath S., Le Quesne J., Tice D.A., Alrifai D., von Karstedt S., Montinaro A., Guppy N., Waller D.A., Nakas A., Good R., Holmes A., Walczak H., Fennell D.A., Garnett M., Iorio F., Wessels L., McDermott U, & Janes S.M. (2018). Loss of functional BAP1 augments sensitivity to TRAIL in cancer cells. eLife, 7, e30224.

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Apoptosis and Cell Death Analysis

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Treated and untreated cells were trypsinized 48 hr after treatment, washed once with 1X PBS and resuspended in 100 μΛ 1X Annexin V binding buffer (BD Biosciences), 4 μL Annexin V AlexaFluor647-conjugated antibody (A23204, ThermoFisher Scientific) and 1.8 μL LIVE/DEAD fixable violet dead cell dye (L34963, ThermoFisher Scientific). After incubation at room temperature for 30 min, 300 μL 1X Annexin V binding buffer was added to each sample. Stained cells were analyzed on a FACSCanto II flow cytometer using FlowJo software.

Walter J., Hertel C., Tannock G.W., Lis C.M., Munro K, & Hammes W.P. (2001). Detection of Lactobacillus, Pediococcus, Leuconostoc, and Weissella Species in Human Feces by Using Group-Specific PCR Primers and Denaturing Gradient Gel Electrophoresis. Applied and Environmental Microbiology, 67(6), 2578-2585.

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