For detection of apoptosis, treated and untreated MSC cells (1×106/plate) were stained with annexin V-conjugated to fluorescein isothiocyanate (FITC) and propidium iodide (PI) using the annexin-V-FITC staining kit (BD Bioscience, San Jose, CA, USA) according to the manufacturer's instructions. Briefly, cells were suspended in 400 µL of annexin V binding buffer and incubated with 3 µL of annexin V and 3 µL of PI for 15 minutes at room temperature in the dark. Images of at least 10,000 cells were acquired in a Beckman Coulter's Navios Tetra system using Kaluza analysis software. Percentages of cells undergoing apoptosis were determined by dual-color analysis. This staining allowed us to distinguish three subsets of cells: viable cells (annexin V negative and PI negative), early apoptotic cells (annexin V positive and PI negative), late apoptotic cells (annexin V positive and PI positive) and necrotic cells (annexin V negative and PI positive). Immediately after staining, the cells were analyzed in a flow cytometer using 488-nm excitation and a 525-nm band pass filter for FITC and a 620-nm filter for PI detection.
Annexin 5 fitc staining kit
Manufactured by BD
Sourced in United States, Germany
The Annexin V-FITC staining kit is a laboratory reagent used for detecting and quantifying apoptosis, a type of programmed cell death. The kit provides Annexin V, a protein that binds to phosphatidylserine, a lipid that is exposed on the surface of cells undergoing apoptosis. The Annexin V is conjugated to the fluorescent dye FITC, allowing for the visualization and analysis of apoptotic cells using flow cytometry or fluorescence microscopy.
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Lab products found in correlation
Cytomics fc500 flow cytometer, by Beckman Coulter (2 mentions)
Facscalibur flow cytometry system, by BD (1 mentions)
Texas red phalloidin, by Thermo Fisher Scientific (1 mentions)
P crkl y207, by Cell Signaling Technology (1 mentions)
Y 27632, by Bio-Techne (1 mentions)
Accuri c6, by BD (1 mentions)
Facscan, by BD (1 mentions)
Facs canto, by Beckman Coulter (1 mentions)
Cellquest software, by BD (1 mentions)
Facscan flow cytometer, by Tree Star (1 mentions)
Tmr red, by Roche (1 mentions)
Cytoflex s flow cytometer, by Beckman Coulter (1 mentions)
Hcs lipidtox green neutral lipid stain, by Thermo Fisher Scientific (1 mentions)
Facscalibur cytometer, by BD (1 mentions)
Acutase, by Innovative Cell Technologies (1 mentions)
Facscalibur system, by BD (1 mentions)
Cytomicstm fc500, by Beckman Coulter (1 mentions)
Notch1 sirna, by Santa Cruz Biotechnology (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
Software, by FlowJo (1 mentions)
29 protocols using annexin 5 fitc staining kit
1
Annexin V-FITC and PI Apoptosis Assay
2
Annexin-V-FITC Flow Cytometry for Apoptosis
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Apoptosis was analyzed by flow cytometry using propidium iodide (PI) and the annexin-V-FITC staining kit (BD Biosciences, Vienna, Austria) following manufacturer's manual.
Data were analyzed by using FlowJo software version 7.2.5 (Tree Star Inc., Ashland, OR, USA).
Data were analyzed by using FlowJo software version 7.2.5 (Tree Star Inc., Ashland, OR, USA).
3
Apoptosis Analysis via Annexin V-FITC
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An Annexin V-FITC staining kit (BD Bioscience, Franklin Lakes, NJ, USA) was used to analyze apoptotic cell death. MCF-7/Dox cells grown in 60 mm diameter dishes were treated with either DOX or DHA alone or in combination for 24 h. After trypsinization and centrifugation, the cell pellets were washed with PBS and suspended in 100 μL of binding buffer containing 5 μL of Annexin V-Fluorescein isothiocyanate (FITC) and 5 μL of propidium iodide (PI). The stained cells were then incubated for 15 min at room temperature and analyzed using a FACSCalibur flow cytometry system (BD Bioscience, Franklin Lakes, NJ, USA).
4
Multiparametric analysis of leukemic cells
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General biochemicals and reagents were obtained from Sigma Aldridge (Sigma Aldridge, Poole, UK) unless otherwise specified. Fluorochrome-conjugated antibodies recognising the following antigens were employed: CD19, CD5, CD23, CD14, CD3, CD4 all from (BD Pharmingen, Oxford, UK). Unconjugated antibodies recognised: CD29 (clone P5D2 Abcam, Cambridge, UK), CRKL (ab32018 Abcam), P-CRKL (Y207) ( #3181 Cell Signaling Technology, New England Biolabs, Hitchin, UK), ABL (ABL clone 8E9 Abcam), ARG (181B11 Abcam). Secondary antibodies used for immunofluorescence were (Goat anti-mouse FITC Ab5999 and goat anti-mouse tr ab6003 Abcam). Phalloidin probes for F-actin were nitrobenzoxadiazole (NBD)-pallacidin (Invitrogen) and Texas Red phalloidin (Invitrogen). Other agents were CXCL12 (Peprotech EC Ltd, London, UK); Y27632 (Tocris Bioscience, Bristol, UK); and imatinib (LKT Laboratories, Alexis Corporation, Lausen, Switzerland). Annexin V FITC Staining kit (BD Pharmingen) was used with manufacturers protocols to determine apoptotic cell death. Immunoperoxidase staining used standard reagents and protocols (DAKO UK Ltd, Ely, UK).
5
Annexin V-FITC Assay for Leishmania Apoptosis
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The phosphatidylserine externalization by the parasites was assessed using an annexin V-FITC staining kit (BD Biosciences, San Diego, USA). Briefly, promastigotes (2×106 cells/mL) were incubated in the absence or presence of LQB-118 (5, 10 or 20 µM) for 24 and 48 h at 28°C. The cells were washed twice and resuspended in 100 µl binding buffer (10 mM HEPES, 150 mM NaCl and 2.5 mM CaCl2), containing 1 µl annexin V-FITC. After 20 min, the cells were washed twice and resuspended in 300 µl of binding buffer; then, at the time of data acquisition, 1.67 µg/mL of propidium iodide was added. The data were acquired using a BD Accuri C6 flow cytometer (BD Accuri, Ann Arbor, MI, USA) and analyzed with CFlow software.
6
Annexin V-FITC Apoptosis Assay
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The Annexin V-FITC staining kit from BD Biosciences (San Jose, CA, USA) was used to determine and quantify the apoptotic cells using flow cytometry, according to the manufacturer’s instruction. In brief, the collected cells were suspended in the supplied binding buffer, and then stained with FITC-conjugated annexin V and PI at room temperature (RT) for 20 min in the dark. The fluorescent intensities of the cells were detected using flow cytometry, and the annexin V+/PI− and annexin V+/PI+ cell populations were considered indicators of apoptotic cells.
7
Annexin V Apoptosis Assay
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Annexin V staining was done using Annexin V-FITC staining kit (BD Biosciences, San Jose, CA, USA) following the manufacturer’s instruction. Briefly, cells treated with MS-5 were washed with PBS and resuspended in 1x binding buffer containing Annexin V and propidium iodide. Fluorescence intensity was measured by flow cytometry.
8
Annexin V-FITC Apoptosis Assay
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The apoptotic cells were detected using an annexin V–FITC staining kit (BD Pharmingen, CA, USA) following the manufacturer's instructions. Briefly, the cells were harvested, washed with phosphate-buffered saline (PBS), and incubated in 500 μL of binding buffer (PH 7.5, 10 mM HEPES, 2.5 mM CaCl2, and 140 mM NaCl) containing annexin V–FITC for 30 min in the dark. After 400 μL of binding buffer was added to stop the reaction, cells were subjected to analysis on a flow cytometer (FACScan, Becton Dickinson, USA).
9
Annexin V-FITC/PI Apoptosis Assay
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For apoptosis assessments, drug-treated HSC-T6s were EDTA treated and pelleted. Cells were resuspended and washed in 200 µL of binding buffer (Annexin V-FITC Staining kit, BD Biosciences) and Annexin-V-FITC/PI stained for 15 min. Samples were resuspended in fresh binding solution and apoptosis rates were assessed on a flow cytometer.
10
Annexin V-FITC and PI Staining
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