Annexin 5 pi kit

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Annexin V/PI kit is a laboratory tool used to assess cell apoptosis. It contains Annexin V, which binds to phosphatidylserine, and propidium iodide (PI), which stains nucleic acids. This combination allows for the differentiation of viable, early apoptotic, and late apoptotic/necrotic cells through flow cytometric analysis.

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Lab products found in correlation

Facscalibur, by BD (2 mentions) Canto 2, by BD (2 mentions) Facscanto 2, by BD (2 mentions) Muse flow cytometer, by Merck Group (2 mentions) Annexin v 7 aad kit, by Thermo Fisher Scientific (1 mentions) Epics xl, by Beckman Coulter (1 mentions) Facscanto 2 flow cytometer, by BD (1 mentions) Facs canto 2 flow cytometer, by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Fetal bovine serum (fbs), by Biowest (1 mentions) Trypsin ethylenediaminetetraacetic acid edta, by Biowest (1 mentions) Anti cd16 32, by Thermo Fisher Scientific (1 mentions) Annexin 5 fitc propidium iodide, by Thermo Fisher Scientific (1 mentions) P akt antibody, by Cell Signaling Technology (1 mentions) Poly adp ribose polymerase parp, by Santa Cruz Biotechnology (1 mentions) Thiostrepton foxm1 selective inhibitor, by Bio-Techne (1 mentions) Bcl2 associated x (bax), by Santa Cruz Biotechnology (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) Foxm1, by Santa Cruz Biotechnology (1 mentions) Caspase 3, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

Annexin V (493 citations) Annexin V/PI (33 citations) Annexin V kit (26 citations)

33 protocols using annexin 5 pi kit

Mycobacterial infection-induced apoptosis in macrophages

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RAW264.7 cells were seeded into 6-well plates, and adherent monolayers were infected with Ms::Vector, Ms::Rv0928, Ms:: Rv1096, Ms:: Rv3033, or Ms:: Rv3369 (MOI = 10) for 48 h. The apoptotic cells were measured by FACS using an Annexin V/PI kit (eBioscience, San Diego, CA, USA). BMDM cells were seeded into 6-well plates, and adherent monolayers were infected with Ms::Vector or Ms:: Rv3033 (MOI = 10) for 48 h. The apoptotic cells were measured by FACS using an Annexin V/PI kit (eBioscience, San Diego, CA, USA). RAW-Vector and RAW-Rv3033 cells were seeded into 6-well plates, and adherent monolayers were infected with BCG (MOI = 10) with or without pretreatment for 1 h with TUDCA (500 μg/ml) for 36 h. The apoptotic cells were measured by FACS using Annexin V-7-AAD kit (eBioscience, San Diego, CA, USA). RAW-Vector and RAW-Rv3033 cells were seeded into 6-well plates, and adherent monolayers were infected with H37Ra for 36 h. The apoptotic cells were measured by FACS using Annexin V/7-AAD kit (eBioscience, San Diego, CA, USA).

Zhang W., Lu Q., Dong Y., Yue Y, & Xiong S. (2018). Rv3033, as an Emerging Anti-apoptosis Factor, Facilitates Mycobacteria Survival via Inhibiting Macrophage Intrinsic Apoptosis. Frontiers in Immunology, 9, 2136.

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Annexin V/PI Apoptosis Assay

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After the designated treatment, cells were washed and labeled with Annexin V/PI kit (eBioscience, 800-8005-72) for 15 min at room temperature in the dark, according to the manufacturer’s instructions. The images were captured using fluorescence microscopy (OLYMPUS, IX81), and the percentage of apoptosis was measured by Flow cytometry (Beckman-Coulter Epics XL).

Yi J., Gong X., Yin X.Y., Wang L., Hou J.X., Chen J., Xie B., Chen G., Wang L.N., Wang X.Y., Wang D.C, & Wei H.L. (2022). Parthenolide and arsenic trioxide co-trigger autophagy-accompanied apoptosis in hepatocellular carcinoma cells. Frontiers in Oncology, 12, 988528.

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Autophagy and Apoptosis Analysis of RAW264.7 Cells Infected with Mycobacteria

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RAW264.7 cells were infected with MS_WT or MS_Rv0790c at a MOI of 10. The cells were then treated with trypsin 4 h post-infection, rinsed with 1x PBS containing 0.05% saponin, and incubated with an anti-LC3 antibody (CST, 3868) for 30 min. Finally, cells were incubated with anti-rabbit IgG488 (CST, 4412) for 30 min. The LC3 positive cells were acquired using a FACS Canto II flow cytometer (Becton Dickinson) and analyzed using FlowJo 7.6.1. For cellular apoptosis detection, RAW264.7 cells were seeded on a 24-well plate and infected with MS_WT or MS_Rv0790c at MOI = 10 for 48 h. Apoptotic cells were measured by a FACS Canto II flow cytometer using an Annexin V/PI kit (eBioscience) and analyzed using FlowJo 7.6.1.

Fang J., Dong C, & Xiong S. (2022). Mycobacterium tuberculosis Rv0790c inhibits the cellular autophagy at its early stage and facilitates mycobacterial survival. Frontiers in Cellular and Infection Microbiology, 12, 1014897.

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Cytotoxicity evaluation of β-elemene

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ß-elemene was purchased from Abcam (Abcam, USA).
Dulbecco’s modified Eagle’s medium (DMEM) and
penicillin/streptomycin solution were procured from
Atocel (Austria). Trypsin-ethylene diamine tetra-acetic
acid (EDTA) and fetal bovine serum (FBS) inactivated
with heat was purchased from Biowest company (France).
3-(4 (link),5 (link)-dimethylthiazol-2-yl)-2,5diphenyltetrazolium
bromide (MTT) and dimethyl sulfoxide (DMSO) were
purchased from Merck (Germany) Annexin V/PI kit was
purchased from Ebioscience company (CA).

Balavandi Z., Neshasteh-Riz A., Koosha F., Eynali S., Hoormand M, & Shahidi M. (2019). The Use of ß-Elemene to Enhance Radio Sensitization of A375 Human Melanoma Cells. Cell Journal (Yakhteh), 21(4), 419-425.

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Apoptosis and TFRC Expression Analysis

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Cell apoptosis were measured by Annexin V/PI kit (eBioscience) according to the manufacturer’s instructions. To detect surface expression of TFRC, isolated BMMs were firstly blocked with anti-CD16/32 (eBioscience) and then stained with PE-conjugated TFRC (eBioscience) in PBS containing 2% FBS on ice for 20 minutes. Then cells were washed with cold PBS, resuspended, and detected by flow cytometry (University of Iowa Flow Cytometry Core, Iowa City, IA).

Gu Z., Wang H., Xia J., Yang Y., Jin Z., Xu H., Shi J., De Domenico I., Tricot G, & Zhan F. (2015). Decreased Ferroportin Promotes Myeloma Cell Growth and Osteoclast Differentiation. Cancer research, 75(11), 2211-2221.

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Lymphocyte Apoptosis Analysis via Flow Cytometry

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After 72 h of incubation period, the apoptosis ratio of lymphocytes was quantified by using the Annexin V/PI kit (eBiosciences, USA) according to the manufacturer’s instructions. Flow cytometric analysis was performed by staining PBMC with anti-CD3 and anti-CD4 antibodies for Annexin V/PI detection.

ZIBANDEH N., GENÇ D., İNANÇ N., DİRESKENELİ H, & AKKOÇ T. (2019). IFN-γ stimulated dental follicle mesenchymal stem cells regulate activated lymphocyte response in rheumatoid arthritis patients in vitro. Turkish Journal of Medical Sciences, 49(6), 1779-1788.

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Annexin V-PI Apoptosis Assay

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The Annexin-V/PI kit (eBioscience, San Diego, CA) was used. Cells were plated at a density of 2×10⁵ cells per 10 cm dish and allowed to attach overnight. The day after plating, vehicle or drug was added to each plate and incubated for 24 h. Cells were harvested and combined with the culture supernatants. Cell pellets were washed with PBS then 1X Binding Buffer, then were resuspended in 1X Binding Buffer at 1-5 × 10⁶ cells/mL; 5 μL of fluorochrome-conjugated Annexin V was added to 100 μL of the cell suspension and incubated for 15 min at room temperature, protected from light. The cells were then washed with 1X Binding Buffer and resuspended in 200 μL of 1X Binding Buffer. Next, cells were stained with 5 μL PI Staining Solution and analyzed by flow cytometry within 4 hours on a FACSCalibur flow cytometer (BD Biosciences). Apoptotic cells were quantified with FlowJo software.

Palisoul M.L., Quinn J.M., Schepers E., Hagemann I.S., Guo L., Reger K., Hagemann A.R., McCourt C.K., Thaker P.H., Powell M.A., Mutch D.G, & Fuh K.C. (2017). Inhibition of the Receptor Tyrosine Kinase AXL Restores Paclitaxel Chemosensitivity in Uterine Serous Cancer. Molecular cancer therapeutics, 16(12), 2881-2891.

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Apoptosis Detection by Flow Cytometry

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Apoptosis was detected with flow cytometry using Annexin V/PI kit (eBioscience; NJ, NY, USA). For this purpose, the cells were incubated with nanoplatforms (4 μg/mL) and nanoplatforms/miR-101 for 90 min in six-well plates. In parallel, the cells incubated with nanoplatforms in the presence or absence of miR-101 were also exposed under laser irradiation (808 nm laser, 1.2 W/cm²) for 10 min. After removing the supernatant, the fresh medium and FBS were added to each well and kept for 24, 48, and 72 h. Finally, the cells were detached from the plate and dyed with Annexin V-FITC/propidium iodide (PI) (based kit protocol, eBioscience) in a dark room. The rates of apoptosis and necrosis were quantified by flow cytometry (FITC detected at 515–545 nm and PI measured at 564–606 nm).

Assali A., Akhavan O., Mottaghitalab F., Adeli M., Dinarvand R., Razzazan S., Arefian E., Soleimani M, & Atyabi F. (2018). Cationic graphene oxide nanoplatform mediates miR-101 delivery to promote apoptosis by regulating autophagy and stress. International Journal of Nanomedicine, 13, 5865-5886.

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Inhibition of FoxM1 and COX-2 in Cancer

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Thiostrepton (FoxM1 selective inhibitor) [56 (link)] was purchased from Tocris Cookson Inc (Ellisville, MO). NS398 (COX-2 inhibitor) was purchased from Caymen chemical company, (Ann Arbor, MI). Antibodies against cleaved caspase-3, Cox-2, AKT and p-AKT antibodies were purchased from Cell Signaling Technologies (Beverly, MA). FoxM1, Bax, Beta-actin, caspase-3 and poly (ADP) ribose polymerase (PARP) antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). MMP-9 antibodies were purchased from Anespec, (San Jose, CA). Annexin V/PI kit was purchased from Molecular Probes (Eugene, OR, USA).

Ahmed M., Hussain A.R., Siraj A.K., Uddin S., Al-Sanea N., Al-Dayel F., Al-Assiri M., Beg S, & Al-Kuraya K.S. (2015). Co-targeting of Cyclooxygenase-2 and FoxM1 is a viable strategy in inducing anticancer effects in colorectal cancer cells. Molecular Cancer, 14, 131.

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MSP-induced Apoptosis Analysis

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TP-4 (FIHHIIGGLFSAGKAIHRLIRRRRR) was from Professor Jyh-Yih Chen’s laboratory at the Academia Sinica Institute. A stock solution of MSPs was prepared in phosphate buffered saline (PBS, pH = 7.2), protected from light, and stored at −20 °C. Annexin V/PI kit was purchased from Molecular Probes, Inc. (Eugene, OR, USA). An In Situ Cell Death Detection Kit (TUNEL) was purchased from Roche Applied Science (Mannheim, Germany). MTT, PI, RNase A, Z-IETD-FMK (caspase-8 inhibitor) and Z-LEHD-FMK (caspase-9 inhibitor) were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA) and dissolved in DMSO or PBS.

Kuo H.M., Tseng C.C., Chen N.F., Tai M.H., Hung H.C., Feng C.W., Cheng S.Y., Huang S.Y., Jean Y.H, & Wen Z.H. (2018). MSP-4, an Antimicrobial Peptide, Induces Apoptosis via Activation of Extrinsic Fas/FasL- and Intrinsic Mitochondria-Mediated Pathways in One Osteosarcoma Cell Line. Marine Drugs, 16(1), 8.

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