Annexin 5 fluorescein isothiocyanate fitc propidium iodide apoptosis kit

Manufactured by Beckman Coulter

Sourced in United States

The Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis Kit is a laboratory reagent used to detect and quantify apoptosis in cell populations. The kit utilizes Annexin V, a calcium-dependent phospholipid-binding protein, conjugated to the fluorescent dye FITC, and the DNA-binding dye PI to identify cells undergoing apoptosis and necrosis, respectively.

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Compusyn software, by ComboSyn (2 mentions) Facscalibur flow cytometer, by BD (1 mentions)

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Annexin V-FITC kit (53 citations) Propidium iodide (50 citations) Annexin V-FITC (37 citations) FITC (15 citations) Annexin V-FITC Apoptosis (5 citations) Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) (3 citations) Annexin-V fluorescein isothiocyanate (FITC) kit (3 citations) Annexin V-fluorescein isothiocyanate (3 citations) Annexin V-fluorescein isothiocyanate (FITC) (2 citations) Annexin V-fluorescein (2 citations)

9 protocols using annexin 5 fluorescein isothiocyanate fitc propidium iodide apoptosis kit

Synergistic Apoptosis Induction in AML

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AML cells were treated with LY2603618 and Roscovitine, alone or in combination, or with LY and ABT-199, alone or in combination, and subjected to flow cytometry analysis to determine drug-induced apoptosis using an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis Kit (Beckman Coulter; Brea, CA, USA), as previously described [41 (link), 43 (link)]. Apoptotic events are presented as percentage of AnnexinV+/PI- and Annexin V+/PI+ ± s.e.m. Experiments with AML cell lines were performed 3 independent times in triplicates, while patient sample experiments were performed once in triplicate due to limited sample. Data are presented as mean ± standard errors from one representative experiment. Patient samples were chosen based on availability of adequate sample for the assay. The extent and direction of antileukemic interaction were determined by calculating the combination index (CI) values using CompuSyn software (Combosyn Inc., Paramus, NJ). CI < 1, CI = 1, and CI > 1 indicate synergistic, additive, and antagonistic effects, respectively [41 (link), 47 (link)].

Zhao J., Niu X., Li X., Edwards H., Wang G., Wang Y., Taub J.W., Lin H, & Ge Y. (2016). Inhibition of CHK1 enhances cell death induced by the Bcl-2-selective inhibitor ABT-199 in acute myeloid leukemia cells. Oncotarget, 7(23), 34785-34799.

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Annexin V-FITC/PI Apoptosis Assay

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Apoptosis was determined using an Annexin V-Fluorescein Isothiocyanate (FITC)/Propidium Iodide (PI) Apoptosis Kit (Beckman Coulter; Brea, CA, USA), as described elsewhere.^{20 (link),21 (link)} The mean percentage (± standard error of mean) of annexinV⁺/PI⁻ (early apoptotic) and annexin V⁺/PI⁺ (late apoptotic and/or dead) cells from one representative experiment is shown.

Li X., Su Y., Madlambayan G., Edwards H., Polin L., Kushner J., Dzinic S.H., White K., Ma J., Knight T., Wang G., Wang Y., Yang J., Taub J.W., Lin H, & Ge Y. (2019). Antileukemic activity and mechanism of action of the novel PI3K and histone deacetylase dual inhibitor CUDC-907 in acute myeloid leukemia. Haematologica, 104(11), 2225-2240.

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Apoptosis Analysis of CuB and SCH772984

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BxPC-3 and HPAC cells were treated with CuB and SCH772984 alone or in combination and subjected to flow cytometry analysis to determine drug-induced cell death using an annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis kit (Beckman Coulter, Brea, CA, USA) and a FACS Calibur flow cytometer (Becton Dickinson, San Jose, CA, USA) as previously described [46 (link), 49 (link)].

Zhou J., Zhao T., Ma L., Liang M., Guo Y.J, & Zhao L.M. (2017). Cucurbitacin B and SCH772984 exhibit synergistic anti-pancreatic cancer activities by suppressing EGFR, PI3K/Akt/mTOR, STAT3 and ERK signaling. Oncotarget, 8(61), 103167-103181.

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Apoptosis Induction in AML Cells

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AML cells were treated with MK-1775, Roscovitine, LY2603618, or the indicated combinations and subjected to flow cytometry analysis to determine drug-induced apoptosis using an annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis Kit (Beckman Coulter; Brea, CA, USA), as previously described [29 (link)],[33 (link)]. Results are expressed as percent of annexin V + cells. Experiments with AML cell lines were performed 3 independent times in triplicates and data presented are from one representative experiment, while patient sample experiments were performed once in triplicates. Data are presented as mean values ± standard errors from one representative experiment. Due to limited sample, only three patient samples were evaluated for MK-1775-induced apoptosis by flow cytometry.

Qi W., Xie C., Li C., Caldwell J.T., Edwards H., Taub J.W., Wang Y., Lin H, & Ge Y. (2014). CHK1 plays a critical role in the anti-leukemic activity of the wee1 inhibitor MK-1775 in acute myeloid leukemia cells. Journal of Hematology & Oncology, 7, 53.

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Cytarabine and Antimycin A Apoptosis Assay

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Primary AML cells from adult patients were treated with either vehicle, 50 or 100 µM cytarabine, 10 nM Antimycin A, or the combination of 50 or 100 µM cytarabine + 10 nM Antimycin A for 4 h. Post treatment, the cells were washed with fresh media (2×) and resuspended in the absence or presence of 10 nM Antimycin A and cultured for an additional 44 h (for a total treatment time of 48 h). The cells were then stained with Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis Kit (Beckman Coulter; Brea, CA, USA). The stage of apoptotic cells (Annexin V-positive, but early or late apoptosis) is identified by the proportion of AnnexinV+/PI− (early apoptosis) and Annexin V+/PI+ (late apoptosis) cells. This experiment was performed once in triplicates (presented as average ± SEM) due to the limited sample availability. Patient samples were selected based on the availability of an adequate number of cells to perform the assay.

Pitre A., Ge Y., Lin W., Wang Y., Fukuda Y., Temirov J., Phillips A.H., Peters J.L., Fan Y., Ma J., Nourse A., Sinha C., Lin H., Kriwacki R., Downing J.R., Gruber T.A., Centonze V.E., Naren A.P., Chen T, & Schuetz J.D. (2017). An unexpected protein interaction promotes drug resistance in leukemia. Nature Communications, 8, 1547.

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Apoptosis Analysis of AML Cells

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AML cells were treated with vehicle, KPT‐330, KPT‐8602, venetoclax and venetoclax plus KPT‐330 or KPT‐8602, and then underwent flow cytometry analysis utilizing the Annexin V‐fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis kit (Beckman Coulter; Brea, CA, USA) to determine the extent of drug‐induced apoptosis, as previously described.²⁸ Annexin V+/PI‐ and Annexin V+/PI+cells represent early apoptotic and late apoptotic (dead) cells respectively. Results are shown as mean percentage of Annexin V‐positive cells ± the standard error of the mean (SEM) of replicates from two independent experiments. Cells treated with an apoptosis inducer purchased from Beyotime Biotechnology (Shanghai, China) were used as the positive control.

Yu H., Wu S., Liu S., Li X., Gai Y., Lin H., Wang Y., Edwards H., Ge Y, & Wang G. (2022). Venetoclax enhances DNA damage induced by XPO1 inhibitors: A novel mechanism underlying the synergistic antileukaemic effect in acute myeloid leukaemia. Journal of Cellular and Molecular Medicine, 26(9), 2646-2657.

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Annexin V-FITC/PI Apoptosis Assay

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Apoptosis was determined using the Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis Kit (Beckman Coulter, Brea, CA, USA), as described.^{21 (link),22 (link)} Mean percentage of AnnexinV+/PI- (early apoptotic) and Annexin V+/PI+ (late apoptotic and/or dead) ± standard error of the mean (SEM) from one representative experiment is shown.

Li X., Su Y., Hege K., Madlambayan G., Edwards H., Knight T., Polin L., Kushner J., Dzinic S.H., White K., Yang J., Miller R., Wang G., Zhao L., Wang Y., Lin H., Taub J.W, & Ge Y. (2020). The HDAC and PI3K dual inhibitor CUDC-907 synergistically enhances the antileukemic activity of venetoclax in preclinical models of acute myeloid leukemia. Haematologica, 106(5), 1262-1277.

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Apoptosis Measurement in Prostate Cancer

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Prostate cancer cells were treated with the indicated drugs. Drug‐induced apoptosis was determined using the Annexin V‐Fluorescein Isothiocyanate (FITC)/Propidium Iodide (PI) Apoptosis Kit (Beckman Coulter) by following the manufacture's protocol. The experiments were repeated at least three times. Mean percentage of AnnexinV+/PI‐ (early apoptotic) and Annexin V+/PI+ (late apoptotic and/or dead) ± standard errors from triplicates of one representative experiment is shown.

Hu C., Xia H., Bai S., Zhao J., Edwards H., Li X., Yang Y., Lyu J., Wang G., Zhan Y., Dong Y, & Ge Y. (2020). CUDC‐907, a novel dual PI3K and HDAC inhibitor, in prostate cancer: Antitumour activity and molecular mechanism of action. Journal of Cellular and Molecular Medicine, 24(13), 7239-7253.

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Apoptosis Induction in AML Cells

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AML cells were treated with VS-5584, SCH772984, Z-VAD-FMK, or ABT-199 alone or in combination for up to 48 h. Drug-induced apoptosis was determined using an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide PI) apoptosis Kit Beckman Coulter; Brea, CA, USA), as previously described [17 (link),22 (link),23 (link)]. For AML cell lines, experiments were performed 2 independent times in triplicates. Patient samples had limited number of cells and so patient sample experiments were performed once in triplicates. Apoptotic events are presented as mean percentage of Annexin V +/PI− and Annexin V+/PI+ ± SEM from one representative experiment. Synergy was determined by calculating the combination index (CI) values using CompuSyn software (Combosyn Inc., Paramus, NJ). CI < 1, CI = 1, and CI > 1 indicate synergistic, additive, and antagonistic effects, respectively.

Su Y., Li X., Ma J., Zhao J., Liu S., Wang G., Edwards H., Taub J.W., Lin H, & Ge Y. (2017). Targeting PI3K, mTOR, ERK, and Bcl-2 signaling network shows superior antileukemic activity against AML ex vivo. Biochemical pharmacology, 148, 13-26.

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