Fitc annexin 5 apoptosis detection kit with 7 aad

Manufactured by BioLegend

Sourced in United States, United Kingdom

The FITC Annexin V Apoptosis Detection Kit with 7-AAD is a laboratory equipment used to detect and quantify apoptosis in cell populations. It utilizes the binding of Annexin V, a protein that has a high affinity for phosphatidylserine, which is exposed on the surface of apoptotic cells. The kit also includes 7-AAD, a dye that stains the DNA of late apoptotic and necrotic cells. This combination allows for the identification and differentiation of viable, early apoptotic, and late apoptotic/necrotic cells through flow cytometry analysis.

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Lab products found in correlation

Facscanto 2, by BD (6 mentions) 7 aad, by BioLegend (6 mentions) Facscanto 2 flow cytometer, by BD (5 mentions) Facsdiva software, by BD (4 mentions) 7 aad, by BD (3 mentions) Fortessa, by BD (3 mentions) Cytofix cytoperm kit, by BD (3 mentions) Ionomycin, by Merck Group (3 mentions) Facscalibur, by BD (3 mentions) Accuri c6 flow cytometer, by BD (2 mentions) Accucheck counting beads, by Thermo Fisher Scientific (2 mentions) Ki67 antibody, by BioLegend (2 mentions) Foxp3 fix perm kit, by BioLegend (2 mentions) Golgiplug, by BD (2 mentions) Anti cd4 pacific blue, by BioLegend (2 mentions) Anti cd8 alexa fluor 700, by BioLegend (2 mentions) Flow cytometry, by BD (2 mentions) Attune nxt flow cytometer, by Thermo Fisher Scientific (2 mentions) Cd11b pe cy7, by Thermo Fisher Scientific (2 mentions) Cd45 bv605, by BioLegend (2 mentions)

Close spelling variants of this product

7-AAD (474 citations) Annexin V (328 citations) Annexin V-FITC (323 citations) FITC Annexin V Apoptosis Detection Kit (118 citations) Apoptosis detection kit (47 citations) FITC Annexin V kit (45 citations) Annexin V Apoptosis Detection Kit (30 citations) Annexin V-FITC Apoptosis Kit (18 citations) FITC Annexin V Apoptosis Detection Kit with 7-aminoactinomycin D (7-AAD) (18 citations) Annexin V Apoptosis Detection Kit with 7-AAD (11 citations)

91 protocols using fitc annexin 5 apoptosis detection kit with 7 aad

Evaluating Anti-α10-SAP Antibody's Effects on Glioblastoma Cells

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The U3054MG GBM cells were seeded at a density of 2500 cells/well in an uncoated 96-well plate, and treated with anti-ctrl-SAP and anti-α10-SAP antibody for 7 d. Whole microscopy images of each well were taken and spheres that had reached a diameter of 100 µm were counted in the digital images using ImageJ software (ImageJ 1.51k, LOCI, University of Wisconsin, Madison, WI, USA) [46 (link)].
The cell viability was determined by using the WST-1 assay (Roche, Mannheim, Germany) according to the manufacturer´s instructions. All conditions were done in triplicate.
To analyze apoptosis, cells were stained with the Annexin V-FITC apoptosis detection kit with 7-AAD (BioLegend, San Diego, CA, USA) according to the supplier’s protocol, and the amount of bound Annexin V-FITC and 7-AAD was quantified with a BD Accuri C6 flow cytometer.

Munksgaard Thorén M., Chmielarska Masoumi K., Krona C., Huang X., Kundu S., Schmidt L., Forsberg-Nilsson K., Floyd Keep M., Englund E., Nelander S., Holmqvist B, & Lundgren-Åkerlund E. (2019). Integrin α10, a Novel Therapeutic Target in Glioblastoma, Regulates Cell Migration, Proliferation, and Survival. Cancers, 11(4), 587.

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Osteoclast Viability Assay

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Cell viability of osteoclasts was analyzed with the Annexin V-FITC Apoptosis Detection Kit with 7-AAD (#640922, Biolegend) according to the manufacturer’s instructions. Fluorescence was recorded on a FACSymphony A1 flow cytometer (BD) and data was analyzed with FlowJo (v10.7, BD).

Chandrabalan S., Dang L., Hansen U., Timmen M., Wehmeyer C., Stange R., Beißbarth T., Binder C., Bleckmann A, & Menck K. (2024). A novel method to efficiently differentiate human osteoclasts from blood-derived monocytes. Biological Procedures Online, 26, 7.

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Measuring Cell Apoptosis with FITC Annexin V

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Assays were performed using the FITC Annexin V Apoptosis Detection Kit with 7-AAD (from BioLegend) and analyzed using a BD FACSAria Fusion instrument controlled by BD FACS Diva software v8.0.1 (BD Biosciences).

Srivastava S., Nataraj N.B., Sekar A., Ghosh S., Bornstein C., Drago-Garcia D., Roth L., Romaniello D., Marrocco I., David E., Gilad Y., Lauriola M., Rotkopf R., Kimchi A., Haga Y., Tsutsumi Y., Mirabeau O., Surdez D., Zinovyev A., Delattre O., Kovar H., Amit I, & Yarden Y. (2019). ETS Proteins Bind with Glucocorticoid Receptors: Relevance for Treatment of Ewing Sarcoma. Cell Reports, 29(1), 104-117.e4.

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Apoptosis Assay in p53-Depleted Cells

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A total of 150,000 cells/well were plated in six-well plates. Approximately 24 h later, cells were treated with either DMSO or 3 μM NMDi for approximately 72 h. Cells were then trypsinized, washed two times with ice-cold phosphate-buffered saline (PBS), and processed for staining using FITC Annexin V Apoptosis Detection Kit with 7AAD (Cat # 640922, BioLegend), according to manufacturer’s instructions. Cells were analyzed by Flow Cytometry.
For assessing apoptosis in p53-depleted A549 cells, cells transfected with either siControl or sip53 (150,000 cells/well) were plated in triplicate in six-well plates and ∼ 24 h later treated with either DMSO or NMDi and processed as above. Experiments were repeated twice.

Gudikote J.P., Cascone T., Poteete A., Sitthideatphaiboon P., Wu Q., Morikawa N., Zhang F., Peng S., Tong P., Li L., Shen L., Nilsson M., Jones P., Sulman E.P., Wang J., Bourdon J.C., Johnson F.M, & Heymach J.V. (2021). Inhibition of nonsense-mediated decay rescues p53β/γ isoform expression and activates the p53 pathway in MDM2-overexpressing and select p53-mutant cancers. The Journal of Biological Chemistry, 297(5), 101163.

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Characterizing Tumor Cell Invasion and Migration

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Basement membrane invasion was determined in matrigel invasion chambers (BD Biosciences, Bedford, MA) (6 (link)). Matrigel migration was measured at 48 hours in triplicate wells and compared with phrGFP vector controls. To evaluate migration via scratch assay, OVCAR3 (0.5 × 10⁶) cells were cultured in a 6-well plate for 24 hrs until 90%−95% confluent. Wound line was created by scratching the plate with a 200 μl micropipette tip. Cells were washed with pH 7.2 phosphate buffered saline (PBS) 2 times to remove floating cells and cultured in RPMI containing 1% FBS for up to 72 hrs. Photos were taken with a microscope at the indicated times. The average width of the gaps was calculated with image j. For spheroid formation, 0.5 × 10⁴ OVCAR3 cells were seeded in the 3D culture qualified 96 well spheroid formation plate (Trevigen). Plates were centrifuged at 200g for 3 min and treated with flavopiridol at 0.25 μM, 0.5 μM, or 1 μM three days later. Spheroids in each well at were photographed at 24 hrs, 48 hrs and 72 hrs. Viability at 72 hrs was assessed via CellTiter-Glo (Promega). Flavopiridol-induced apoptosis was evaluated using FITC Annexin V Apoptosis Detection Kit with 7-AAD (Biolegend) according to the manufacturer’s instructions and then analyzed by flow cytometry (BD Biosciences).

Rao T.D., Xu M., Eng S., Yang G., Manson R., Rosales N., Kumar R., Veillard I.E., Zhou Q., Iasonos A., Ouerfelli O., Djaballah H., Spriggs D.R, & Yeku O.O. (2022). Dual Fluorescence Isogenic Synthetic Lethal Kinase Screen and High-Content Secondary Screening for MUC16/CA125 Selective Agents. Molecular cancer therapeutics, 21(5), 775-785.

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Evaluating Integrin-Mediated Apoptosis in Glioma Cells

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Example 9

Cells are dependent on integrin adhesion to the extracellular matrix in order to survive. If these integrin-matrix bonds are broken, cells may go into apoptosis. In order to elucidate whether induction of apoptosis occurs in glioma (GBM) cells when integrin α10 bonds are broken, apoptosis assays are performed.

Glioma cells (GBM) are cultured in 6-well plates and incubated overnight for determination of apoptosis. Unconjugated or ADC monoclonal antibodies against integrin alpha 10 subunit, or IgG control antibodies are added the day after. Morphological changes are observed using an inverted microscope, and cells are harvested for flow cytometry analysis after 72 h treatments to observe early and late apoptosis. The degree of apoptosis induced by integrin alpha 10 subunit antibody is determined by a commercially available apoptosis detection kit such as FITC annexin V Apoptosis detection kit with 7-AAD (Biolegend) and the data is analyzed by flow cytometry. In flow cytometry analysis, Annexin V-FITC single positive cells represent early apoptotic cells. Annexin V-FITC and 7-AAD (7-amino-actinomycin D) double positive staining cells represent late apoptotic cells. Positive staining for 7-AAD only represents the necrotic population of cells.

Glioma cells incubated with an antibody against integrin alpha 10 subunit show enhanced apoptosis.

US10994022B2. Detection and treatment of malignant tumours in the CNS (2021-05-04). Xintela AB [SE]. Inventors: Evy Lundgren-Åkerlund [SE], Ramiro Gisler [SE], Jan Talts [SE].

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Comprehensive Splenic T Cell Analysis

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Spleens were harvested after mice were sacrificed 48 hours after CLP, and then were processed to single-cell suspensions though a 70 μm filter. Splenocytes were rinsed with 10 mL cold PBS, and 200 μL from each spleen was put into a 96-well plate for staining. Anti-CD3 (BD, clone 500A2), anti-CD4 (clone RM4-5, BioLegend), anti-CD8 (clone MCD0830, Invitrogen), anti-CD44 ( clone IM7, BioLegend), anti-CD226 (clone 10E5, BioLegend), anti-PD-1 (clone 29F.1A12, BioLegend), anti-2B4 (clone eBio244F4, Thermo Fisher Scientific), and anti-TIM-3 (clone RMT3-23, BioLegend) were used for surface staining to determine T cell phenotype. For the detection of cell apoptosis, splenocytes were stained with a FITC Annexin V apoptosis detection kit with 7-AAD (BioLegend). Tregs were identified via intracellular staining for Foxp3-APC (clone FJK-16S, eBioscience). Splenocytes were surface stained for anti-CD62L (MEL-14), anti-CD69 (H1.2F3), and then permeabilized using a Foxp3 kit (BD Biosciences) and stained with anti-Foxp3, anti-CTLA-4 (UC10-489), and anti-Helios (22F6, all Abs from BioLegend). AccuCheck Counting Beads (Thermo Fisher Scientific) were added after staining to calculate the absolute number of cells per spleen.

Sun Y., Anyalebechi J.C., Sun H., Yumoto T., Xue M., Liu D., Liang Z., Coopersmith C.M, & Ford M.L. (2021). Anti-TIGIT differentially affects sepsis survival in immunologically experienced versus previously naive hosts. JCI Insight, 6(5), e141245.

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Liver Cancer Apoptosis Induction

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The reagents and kits commonly used in this study induclude doxorubicin (Solarbio, USA), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (Yeyuan Biotech, Shanghai), Fe₃O₄-SH (Ruixi Biotech, Shanxi), N-hydroxysuccinimide (Yeyuan Biotech, Shanghai), Anti-LGMN antibody (ABclonal, USA), DMEM medium (Gibco, USA), fetal bovine serum (Gibco, USA), FITC Annexin V Apoptosis Detection Kit with 7-AAD (BioLegend, USA), and TUNEL Apoptosis Kit (RuiBo, Shanghai). The PGMLV-CMV-H_LGMN-3 × Flag-EF1-ZsGreen1-T2A-Puro plasmid was constructed by Jiman Biological Company (Shanghai, CN). Human liver cancer cells (PLCs) were kindly provided by National Liver Cancer Science Center of China (Shanghai, CN). BABL/c male nude mice were purchased from the Animal Experimental Center of Naval Military Medical University (Shanghai, CN). All animal procedures were performed in accordance with the Guidelines for Care and Use of Laboratory Animals of “Naval Military Medical” University and approved by the Animal Ethics Committee of “Naval Military Medical.

Li J., Li L., Lv Y., Zou H., Wei Y., Nie F., Duan W., Sedike M., Xiao L, & Wang M. (2020). The construction of the novel magnetic prodrug Fe3O4@DOX and its antagonistic effects on hepatocarcinoma with low toxicity. RSC Advances, 10(48), 28965-28974.

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Quantitative Analysis of Cellular Apoptosis and Necrosis

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Following the treatment of desired drugs at indicated concentrations, cell viability measured by MTT assay, cell detachment assay by counting the detached (floating) and attached (collected by trypsinization) cells were performed as previously described [22 (link)]. Flow cytometry was used for quantitative analysis of apoptotic and necrotic cells as previously described [22 (link)], using a FITC Annexin V Apoptosis Detection Kit with 7-AAD (BioLegend).
Cytation 3 (BioTek Instruments), a cell-based multi-mode microplate reader and image analysis system, was used for quantitative analysis of Sytox Green (Life Technologies) uptake by necrotic cells. The ratio of necrotic cells with compromised cellular membranes can be measured by fluorescence imaging of 24-well plates combined with image-based quantitative analyses of green nuclei (Sytox Green positive cells) and blue nuclei (Hoechst 33342 staining for total cells). The samples were prepared in duplicate and at least 5,000 cells were analyzed for each well. At least five independent experiments were analyzed.

Wei L., Surma M., Gough G., Shi S., Lambert-Cheatham N., Chang J, & Shi J. (2015). Dissecting the Mechanisms of Doxorubicin and Oxidative Stress-Induced Cytotoxicity: The Involvement of Actin Cytoskeleton and ROCK1. PLoS ONE, 10(7), e0131763.

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Apoptosis Quantification in Tumor Cells

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Apoptosis was measured according to the protocol described for FITC Annexin V Apoptosis Detection Kit with 7-AAD (BioLegend). Briefly, the tumor cells were labeled by CellTrace™ Violet dye, and cocultured with DC-CIK cells, ipilimumab, or hIgG1 treated DC-CIK cells at an E/T ratio of 5:1. After incubation with 10 μg/ml G28.5 or mIgG1 control for 10-14 h, all cells were collected and resuspended in 100 μl Annexin V binding buffer. Five microliter FITC Annexin V and 7-AAD were added and incubated at room temperature for 15 min in the dark. Another 200 μl Annexin V binding buffer was added to each tube, and samples were then analyzed by flow cytometry.

Zhang Y., Wu X., Sharma A., Weiher H., Schmid M., Kristiansen G, & Schmidt-Wolf I.G. (2022). Anti-CD40 predominates over anti-CTLA-4 to provide enhanced antitumor response of DC-CIK cells in renal cell carcinoma. Frontiers in Immunology, 13, 925633.

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