Annexin 5 apc 7 aad apoptosis detection kit

Manufactured by BioLegend

Sourced in United States

The Annexin V-APC/7-AAD apoptosis detection kit is a laboratory tool used to identify and quantify apoptotic cells. It contains Annexin V conjugated with the fluorescent dye APC and the nuclear stain 7-AAD. This combination allows for the detection of cells in early and late stages of apoptosis.

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Lab products found in correlation

Trypsin, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions) Annexin 5 fitc pi apoptosis detection kit, by BD (1 mentions) Facs flow cytometer, by BD (1 mentions) Trypsin ethylenediaminetetraacetic acid edta, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Ham s f 12k medium, by Merck Group (1 mentions) 3 4 5 dimethylthiazol 2 yl 2 5 diphenyl tetrazolium bromide mtt hoechst 33258, by Merck Group (1 mentions)

Spelling variants of this product

Annexin V-APC (225 citations) APC Annexin V Apoptosis Detection Kit (61 citations) Apoptosis detection kit (47 citations) Annexin V Apoptosis Detection Kit (30 citations) APC Annexin V kit (5 citations) Annexin V detection kit (5 citations) Annexin-V-APC/7-AAD Kit (4 citations) APC Annexin V apoptosis kit (3 citations) Annexin V-APC/7-AAD apoptosis kit (2 citations) Annexin V-APC/7-AAD (2 citations)

6 protocols using annexin 5 apc 7 aad apoptosis detection kit

Cell Viability and Apoptosis Analysis

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After transfection for 48 h, we tested cell viability using CCK-8 assay (n=3/group). CCK-8 buffer (MCE, Shanghai, China) (10 µL) was added into the wells and further cultured in an incubator at 37 °C for 4 h. The microplate reader was used to measure the absorbance value at 450 nm.
The apoptosis analyze was implemented after 48 h transfection (n=3/group). After digestion with 0.25% ethylenediaminetetraacetic acid-free trypsin (Invitrogen, Carlsbad, CA, USA) and washing, the cells were centrifuged at 1,500 rpm to remove the supernatant. Apoptosis was evaluated using an Annexin V-APC/7-AAD apoptosis detection kit (BioLegend, San Diego, CA, USA).

Zhu H., Yue H., Xie Y., Chen B., Zhou Y, & Liu W. (2021). Bioinformatics and integrated analyses of prognosis-associated key genes in lung adenocarcinoma. Journal of Thoracic Disease, 13(2), 1172-1186.

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Apoptosis Detection by Annexin V-APC/7-AAD Flow Cytometry

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Cell apoptosis was measured with an Annexin V APC/7-AAD apoptosis detection kit (Biolegend, USA). Cells were harvested by trypsinization without EDTA and suspended in 1 × binding buffer at a density of 5.0 × 10⁶cells/ml. Then, 5 μL of APC-conjugated Annexin V and 5 μL of 7-AAD were added to the cell suspension per 100 μL and incubated for 15 min in the dark at room temperature. Then, Annexin V Binding Buffer was used to stop the reaction. The samples were analyzed by flow cytometry (Beckman-Coulter, China).

Liu M., Zhang D., Zhou X., Duan J., Hu Y., Zhang W., Liu Q., Xu B, & Zhang A. (2022). Cell-free fat extract improves ovarian function and fertility in mice with premature ovarian insufficiency. Stem Cell Research & Therapy, 13, 320.

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Apoptosis Analysis by Flow Cytometry

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Flow cytometry was performed to analyze apoptosis rate. Treated cells were digested and resuspended as a single-cell suspension and then washed twice in PBS. Apoptosis was measured on the same FACS Canto II (BD) according to the manufacturer instruction of Annexin V-FITC/PI Apoptosis Detection kit (BD) and Annexin V-APC/7-AAD Apoptosis Detection kit (BioLegend, San Diego, CA, USA).

Jing C., Li X., Zhou M., Zhang S., Lai Q., Liu D., Ye B., Li L., Wu Y., Li H., Yue K., Chen P., Yao X., Wu Y., Duan Y, & Wang X. (2021). The PSMD14 inhibitor Thiolutin as a novel therapeutic approach for esophageal squamous cell carcinoma through facilitating SNAIL degradation. Theranostics, 11(12), 5847-5862.

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Cell Viability and Apoptosis Assay

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Cells transduced with the lentiviruses expressing specific shRNAs were analyzed with the trypan blue exclusion assay to determine cell viability (11 (link)). Apoptotic cells were examined with Annexin V/7-AAD staining using the AnnexinV-APC/7-AAD Apoptosis Detection kit (Biolegend, USA). Flow cytometry analysis was performed on a FACS flow cytometer (Becton-Dickinson). Data were analyzed by Flowjo software.

Zhang H., Chen L., Cai S.H, & Cheng H. (2016). Identification of TBK1 and IKKε, the non-canonical IκB kinases, as crucial pro-survival factors in HTLV-1-transformed T lymphocytes. Leukemia research, 46, 37-44.

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MoS2 Nanoparticle Functionalization and Cytotoxicity

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MoS₂ crystal was purchased from Muke Nano Science and Technology Co., Ltd. (Nanjing, China). HS-PEG-NH₂ (molecular weight (MW): 5000) was provided by Xi'an Ruixi biological Co., Ltd. (Xi'an, China). 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT), Hoechst 33,258, and Ham’s F12K medium were obtained from Sigma-Aldrich (St. Louis, USA). Trypsin-ethylenediaminetetraacetic acid (EDTA) and fetal bovine serum (FBS) were purchased from Gibco-BRL (Burlington, Canada). An Annexin V-APC/7-AAD apoptosis detection kit was supplied by Biolegend Company (San Diego, USA). All other reagents were purchased from J&K Scientific, Ltd., and used without further purification.

Chen Z., Wei X., Zheng Y., Zhang Z., Gu W., Liao W., Zhang H., Wang X., Liu J., Li H, & Xu W. (2023). Targeted co-delivery of curcumin and erlotinib by MoS2 nanosheets for the combination of synergetic chemotherapy and photothermal therapy of lung cancer. Journal of Nanobiotechnology, 21, 333.

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Apoptosis Detection in T Cells and Tumor Cells

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Apoptotic cells were detected by the apoptosis assay using flow cytometry. For T cell apoptosis, anti-CD3-pre-activated T cells were co-cultured with tumor cells transfected with ILT4 knockdown lentivirus or pretreated with 0.5 μg/mL control IgG/anti-ILT4/anti-PD-L1/both antibodies for 8 h at a ratio of 2:1. The T cells were then purified with PE-anti-CD3 and apoptotic cells were detected by the Annexin V-APC/7-AAD Apoptosis Detection kit (BioLegend; Cat No. 640930). For tumor cell apoptosis, cell lines transfected with ILT4 knockdown lentiviruses were seeded into 6-well plates and cultured for 48 h. Then cells were harvested and the apoptotic rate was detected. The Annexin V-APC-positive cells (either 7-AAD-negative or -positive) were defined as apoptotic cells.

Chen X., Gao A., Zhang F., Yang Z., Wang S., Fang Y., Li J., Wang J., Shi W., Wang L., Zheng Y, & Sun Y. (2021). ILT4 inhibition prevents TAM- and dysfunctional T cell-mediated immunosuppression and enhances the efficacy of anti-PD-L1 therapy in NSCLC with EGFR activation. Theranostics, 11(7), 3392-3416.

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