Fitc conjugated annexin 5

Manufactured by Merck Group

Sourced in United States

FITC-conjugated annexin V is a fluorescent dye-labeled protein that binds to phosphatidylserine, a phospholipid exposed on the surface of apoptotic cells. It is a tool used in research applications to detect and quantify cells undergoing programmed cell death.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Annexin V-FITC (217 citations) Annexin V (119 citations) Fluorescein isothiocyanate (FITC) conjugated Annexin V (3 citations) FITC)-conjugated Annexin V and PI (Annexin V-FITC Apoptosis Detection Kit, (2 citations)

13 protocols using fitc conjugated annexin 5

Annexin V-PI Apoptosis Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

For the assay, cells (10⁵ per well) were seeded in 24-well plates (Corning), incubated for 24 h, and N-acyl dopamines (5 μM) were added for the next 5 h. Then stromal cells were harvested with trypsin-EDTA solution, washed twice with cold PBS (800 g for 5 min), and the pellet was resuspended in binding buffer (10 mM HEPES/NaOH pH 7.4, 140 mM NaCl, 2.5 mM CaCl₂). Cell suspension (100 μL) was transferred to a 5 mL culture tube and incubated with 5 μL of FITC-conjugated annexin V (1 mg/mL, Sigma-Aldrich, USA) and 5 μL of propidium iodide (PI, 2 mg/mL, Sigma-Aldrich) for 15 min at room temperature in the dark. A total of 400 μL of the binding buffer was added to each sample tube, and cell counts were performed by FACSCalibur flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA). The distribution of fluorescent dyes was analyzed using Cell Quest Research Software (BD, Biosciences). Cells were classified as live (Annexin V−, PI−), necrotic (Annexin V−, PI+), early apoptotic (Annexin V+, PI−), and late apoptotic (Annexin V+, PI+).

Gamisonia A.M., Yushina M.N., Fedorova-Gogolina I.A., Akimov M.G., Eldarov C.M., Pavlovich S.V., Bezuglov V.V., Gretskaya N.M., Sukhikh G.T, & Bobrov M.Y. (2021). N-Acyl Dopamines Induce Apoptosis in Endometrial Stromal Cells from Patients with Endometriosis. International Journal of Molecular Sciences, 22(19), 10648.

+ Open protocol

+ Expand

Quantifying Apoptosis via Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols

At 24 h after irradiation, the cells were harvested and incubated with 5 µL of FITC-conjugated Annexin V and 5 µL of propidium iodide for 15 min (Sigma) according to manufacturer's instructions at room temperature in the dark. The proportions of apoptotic cells were quantified using a FACS Calibur flow cytometer and Cellquest software (BD Biosciences, USA).

Chi Y.F., Qin J.J., Li Z., Ge Q, & Zeng W.H. (2020). Enhanced anti-tumor efficacy of 5-aminolevulinic acid-gold nanoparticles-mediated photodynamic therapy in cutaneous squamous cell carcinoma cells. Brazilian Journal of Medical and Biological Research, 53(5), e8457.

+ Open protocol

+ Expand

Apoptosis and Autophagy Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

Propidium iodide, rapamycin, camptothecin, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), hydrogen peroxide, 4’,6-diamidino-2-phenylindole (DAPI), staurosporine, 2′,7’-dichlorofluorescein diacetate (DCFH-DA), dimethyl sulphoxide (DMSO), FITC-conjugated annexin V, acridine orange, Tris-buffered saline with 0.1% Tween^® 20 detergent and N-acetylcysteine came from Sigma-Aldrich, Merck (St. Louis, MO, USA). The required primary antibodies (cyclin D1, β-actin, p27, p21, MAPLC3I/II, SQSTM1/P62, PARP1 and caspase-3) were from Cell Signaling Technology (Leiden, The Netherlands). The enzyme-labelled mouse and rabbit IgG secondary antibodies were procured from Santa Cruz Biotechnology (Dallas, TX, USA).

Alnuqaydan A.M, & Rah B. (2021). Tamarix articulata Inhibits Cell Proliferation, Promotes Cell Death Mechanisms and Triggers G0/G1 Cell Cycle Arrest in Hepatocellular Carcinoma Cells. Food Technology and Biotechnology, 59(2), 162-173.

+ Open protocol

+ Expand

Measuring Cell Death by Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols

Induction of cell death was analyzed by flow cytometry using FITC-conjugated annexin V (BioVision, USA) and propidium iodide (PI, Fluka) double staining. SK-BR-3, T47D, MDA-MB468 and SW480 cells were seeded into 12-well plates in a density of 5–13 × 10⁴ cells per well in complete medium and allowed to settle for 24 h. The cells were exposed to test compounds in different concentrations for 48 h at 37 °C. Merbarone (Sigma-Aldrich) was used as a positive control at a concentration of 160 µM. After incubation, cells were gently trypsinized, washed with PBS, and suspended with FITC-conjugated annexin V (0.25 μg/mL) and PI (1 μg/mL) in binding buffer (10 mM HEPES/NaOH pH 7.4, 140 mM NaCl, 2.5 mM CaCl₂) at 37 °C for 15 min. Stained cells were analyzed with a Guava 8HT EasyCyte flow cytometer (Millipore) using InCyte software and analyzed by FlowJo software. Results are based on three independent experiments.

Legina M.S., Nogueira J.J., Kandioller W., Jakupec M.A., González L, & Keppler B.K. (2020). Biological evaluation of novel thiomaltol-based organometallic complexes as topoisomerase IIα inhibitors. Journal of Biological Inorganic Chemistry, 25(3), 451-465.

+ Open protocol

+ Expand

Membrane Lipid Visualization in GBM Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The presence and distribution of phosphatidylserine and cholesterol residues on GBM cells plasma membrane were analyzed with a fluorescence microscope Eclipse 80i (Nikon, Kawasaki, Kanagawa Prefecture, Japan). Phosphatidylserine residues were labelled for 10 min at 37 °C with 0.5 mg/mL of FITC-conjugated annexin-V (#A9210; Sigma-Aldrich, St. Louis, MO, USA) in the complete culture medium of cells grown on coverslips (1 × 10⁵ cells/13 mm diameter coverslip) after two extensive rinsings with 0.2M of PBS (pH 7.4).
Cholesterol was labeled for 2 h at 37 °C with a 0.05-mg filipin (#70440; Cayman Chemical, Milan, Italy)/mL of PBS (0.2 M), pH 7.4, in complete culture medium of cells grown on coverslips (1 × 10⁵ cells/13 mm diameter coverslip) after two extensive rinsing with 0.2M of PBS (pH 7.4).
Before fluorescence microscopy, observation slides were extensively rinsed with 0.2M of PBS (pH 7.4) and glycerine mounted medium.

Panzarini E., Tacconi S., Carata E., Mariano S., Tata A.M, & Dini L. (2020). Molecular Characterization of Temozolomide-Treated and Non Temozolomide-Treated Glioblastoma Cells Released Extracellular Vesicles and Their Role in the Macrophage Response. International Journal of Molecular Sciences, 21(21), 8353.

+ Open protocol

+ Expand

Annexin-V/PI Cell Viability Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell viability was assessed with flow cytometry recordings as described [47 (link)]. Briefly, cells were stained with FITC-conjugated Annexin-V (Boehringer Mannheim) and propidium iodide (PI, 1 μg/ml; Sigma), to determine phosphatidyl-serine exposure on the cell surface (increased FITC-conjugated Annexin-V staining) and loss of plasma membrane integrity (PI permeability and staining). Cells were incubated at 37°C in 135 mM NaCl, 10 mM HEPES, 5 mM CaCl₂ and then analyzed on a FACS Canto II flow cytometer (Becton Dickinson). Data acquisition and analysis were performed using FACSDiva software.

Guzzo G., Sciacovelli M., Bernardi P, & Rasola A. (2014). Inhibition of succinate dehydrogenase by the mitochondrial chaperone TRAP1 has anti-oxidant and anti-apoptotic effects on tumor cells. Oncotarget, 5(23), 11897-11908.

+ Open protocol

+ Expand

Apoptosis Induction by UV Irradiation

Check if the same lab product or an alternative is used in the 5 most similar protocols

HT-29, T47D, HS578T, MCF7, HFF-1 and HMEC-1 were induced apoptosis by UV irradiation in 35 mm Petri dishes in the absence of cell culture medium. Specifically, after removal of culture medium, the dishes were placed under a UV lamp/cross-linker (model XL-1000, Spectronics Corporation, New York) and irradiated for various time periods, all within 200 seconds, to achieve induction of cell apoptosis starting at 2–4 h post-UV, but not necrosis or instant death (the total energy strength for different cell types are listed in Table 1). After UV irradiation, the culture medium was added back to the dishes and the cells were cultured for various time periods until analysis. Cell apoptosis was assessed by labeling with FITC-conjugated Annexin V (Sigma) for cell surface PS and YO-PRO-1 iodide (Life Technologies) for cell nuclei. Cell necrosis or dead cells were detected by staining with propidium iodide (PI). Cell apoptosis was also assessed by Western blot to detect cleavage of Poly ADP-Ribose Polymerase (PARP) using a rabbit polyclonal anti-PARP antibody (Roche).

Lv Z., Bian Z., Shi L., Niu S., Ha B., Tremblay A., Li L., Zhang X., Paluszynski J., Liu M., Zen K, & Liu Y. (2015). Loss of cell surface CD47 ‘clustering’ formation and binding avidity to SIRPα facilitate apoptotic cell clearance by macrophage. Journal of immunology (Baltimore, Md. : 1950), 195(2), 661-671.

+ Open protocol

+ Expand

Ephrin-B2 Signaling Regulates Apoptosis

Check if the same lab product or an alternative is used in the 5 most similar protocols

SAS-L1 cells were transiently transfected with EFNB2 siRNA or control scrambled siRNA and cultured for 24 h. Next, the cells were incubated with 2 μg/mL clustered Fc, ephrin-B2/Fc or Eph-B4/Fc. At indicated time points, cells were stained with propidium iodide and FITC-conjugated annexin V (Sigma-Aldrich, St. Louis, MO, USA) and analyzed on a FACScan cytometer using CELLQUEST (Becton Dickinson, San Jose, CA, USA).

Sasabe E., Tomomura A., Tomita R., Sento S., Kitamura N, & Yamamoto T. (2017). Ephrin-B2 reverse signaling regulates progression and lymph node metastasis of oral squamous cell carcinoma. PLoS ONE, 12(11), e0188965.

+ Open protocol

+ Expand

Apoptosis Analysis of CQ and Zinc Treated Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The CNE-2s cells were placed in 6-well plates were starved for 6 h and treated with 1μM CQ and 10μM zinc for 24 h. The cells were harvested, washed twice with cold PBS, and stained with FITC-conjugated annexin V for 20 min and propidium iodide for 5 min (Sigma-Aldrich) in the dark. The stained cells were assessed by flow cytometry (FACSAria ^TM[Ⅲ], BD, Mountain View, USA)), and analyzed by FlowJo vX.0.7 software. The intracellular apoptosis rates were measured by flow cytometry as previously described 20 (link). Briefly, 510⁵ cells were seeded in 6-well plates and then treated as indicated. The cells were washed twice with cold PBS and collected for fluorescence analysis using a flow cytometer (FACSAria TM[Ⅲ], BD).

Ke Y., Wu C., Zeng Y., Chen M., Li Y., Xie C., Zhou Y., Zhong Y, & Yu H. (2020). Radiosensitization of Clioquinol Combined with Zinc in the Nasopharyngeal Cancer Stem-like Cells by Inhibiting Autophagy in Vitro and in Vivo. International Journal of Biological Sciences, 16(5), 777-789.

+ Open protocol

+ Expand

Cell Viability and Apoptosis Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

For the viability assays, cells were seeded in 96-well plates (2×10⁴ cells/well) in complete medium, and incubated overnight at 37°C with increasing concentrations of either BMTP-78 or an admixture of WIFPWIQL and _D(KLAKLAK)₂ at equimolar concentrations (negative control). Viability was determined by measuring the cytoplasmic lactate dehydrogenase (LDH) enzymatic activity with DHL™ Cell Viability & Proliferation Assay Kit (AnaSpec). For the apoptosis assays, OCI-AML cells were seeded in 6-well plates (2 × 10⁵ cells/well), and incubated overnight at 37°C with 20 μM of either BMTP-78 or the control admixture, followed by staining with FITC-conjugated annexin V and propidium iodide (PI, Sigma), and flow cytometry with a BD FACS Canto II.

Staquicini D.I., D’Angelo S., Ferrara F., Karjalainen K., Sharma G., Smith T.L., Tarleton C.A., Jaalouk D.E., Kuniyasu A., Baze W.B., Chaffee B.K., Hanley P.W., Barnhart K.F., Koivunen E., Marchiò S., Sidman R.L., Cortes J.E., Kantarjian H.M., Arap W, & Pasqualini R. (2017). Therapeutic targeting of membrane-associated GRP78 in leukemia and lymphoma: preclinical efficacy in vitro and formal toxicity study of BMTP-78 in rodents and primates. The pharmacogenomics journal, 18(3), 436-443.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!