Annexin 5 fitc pi double stain assay

Manufactured by BD

Sourced in United States

Annexin V FITC/PI double stain assay is a laboratory technique used to detect and quantify apoptosis, a process of programmed cell death. The assay utilizes Annexin V, a calcium-dependent phospholipid-binding protein that has a high affinity for phosphatidylserine, which is exposed on the surface of apoptotic cells. The assay also incorporates propidium iodide (PI), a DNA-binding dye that can penetrate the compromised membranes of late apoptotic or necrotic cells. This combination of Annexin V and PI allows for the identification and differentiation of viable, early apoptotic, late apoptotic, and necrotic cells.

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Lab products found in correlation

Cell counting kit 8 (cck8), by Dojindo Laboratories (2 mentions) In situ cell death detection kit, by Roche (1 mentions) Apoptosis assay kit, by Beyotime (1 mentions) Propidium iodide, by BD (1 mentions)

Close spelling variants of this product

PI/Annexin V stain Annexin V-FITC/PI Annexin V/PI Annexin/PI assay Annexin V stain Annexin V-FITC Annexin V (Annexin V-FITC) Annexin V assay PI) stain Annexin V

5 protocols using annexin 5 fitc pi double stain assay

Annexin V-FITC/PI Flow Cytometry

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Apoptosis was examined by flow cytometric analysis. An Annexin V–FITC/PI double stain assay (BD Biosciences, San Jose, CA, USA) was performed following the manufacturer’s protocol. The analysis was performed with FlowJo software (Treestar, Inc., San Carlos, CA, USA). All the assays were performed in triplicate.

Li Y., Chen F., Chu J., Wu C., Li Y., Li H, & Ma H. (2019). miR-148-3p Inhibits Growth of Glioblastoma Targeting DNA Methyltransferase-1 (DNMT1). Oncology Research, 27(8), 911-921.

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Cell Proliferation and Apoptosis Assay

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Cell proliferation was assessed by Cell Counting Kit-8 (Dojindo, Japan) assay. Cells were seeded at 2000 cells/well into 96-well plates with 100 μL culture medium. The 10 μL of CCK-8 solution was added to the cells at specific time points and cells were incubated for 2 hr at 37°C. The reaction product was quantified according to the manufacturer’s instructions.
Apoptosis was examined by flow cytometric analysis. An Annexin V FITC/PI double stain assay (BD Biosciences, San Jose, CA) was performed following the manufacturer’s protocol. When the cells were treated with Doxorubicin, we performed FVS510 and Annexin V-FITC double staining to avoid the fluorescent signal of the drug. Tumor cell apoptosis in the xenograft tumor tissues was detected by terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling (TUNEL) technology using the in Situ Cell Death Detection Kit (Roche Molecular Biochemicals, Mannheim, Germany). The negative control was incubated with label solution (without terminal transferase) instead of the TUNEL reaction mixture.

Yu T., Guo F., Yu Y., Sun T., Ma D., Han J., Qian Y., Kryczek I., Sun D., Nagarsheth N., Chen Y., Chen H., Hong J., Zou W, & Fang J.Y. (2017). Fusobacterium nucleatum Promotes Chemoresistance to Colorectal Cancer by Modulating Autophagy. Cell, 170(3), 548-563.e16.

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Quantifying Cell Apoptosis with Flow Cytometry

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The cell apoptosis ratio was determined by flow cytometric analysis with an Annexin V FITC/PI double stain assay (BD Biosciences, USA) according to the manufacturer’s instructions. Levels of apoptosis in the xenograft tumor tissues were determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) technology using an Apoptosis Assay Kit (Beyotime, China).

Zhang S., Yang Y., Weng W., Guo B., Cai G., Ma Y, & Cai S. (2019). Fusobacterium nucleatum promotes chemoresistance to 5-fluorouracil by upregulation of BIRC3 expression in colorectal cancer. Journal of Experimental & Clinical Cancer Research : CR, 38, 14.

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Quantifying Apoptosis by Flow Cytometry

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Apoptosis was examined by flow cytometric analysis. An Annexin V-FITC/PI double stain assay (BD Biosciences, San Jose, CA) was performed following the manufacturer’s protocol. The analysis was performed with FlowJo software (Treestar, Inc., San Carlos, CA). All the assays were performed in triplicate apoptosis analyses.

Wang M., Han D., Yuan Z., Hu H., Zhao Z., Yang R., Jin Y., Zou C., Chen Y., Wang G., Gao X, & Wang X. (2018). Long non-coding RNA H19 confers 5-Fu resistance in colorectal cancer by promoting SIRT1-mediated autophagy. Cell Death & Disease, 9(12), 1149.

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Assessing Cell Viability and Apoptosis

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The viability of cells was assessed by Cell Counting Kit-8 (CCK8, Dojindo Molecular Technologies, Japan) according to the manufacturer's instructions. Tumor cells were seeded onto the 96-well plates at a density of 3 × 10³ cells per well. CCK8 solution was added to each
well at the time points of 24 h, 48 h, 72 h, and 96 h. The plate was examined at 450 nm absorption with a microplate reader after incubating for 2 hours at 37°C.
Cell cycles were examined using propidium iodide (PI) (BD Biosciences). Cells were washed with cold PBS and fixed in 75% ethanol −20°C overnight, and then incubated with PI for 15 minutes before flow cytometric analysis. For measuring the extent of apoptosis, CRC cells were analyzed through flow cytometry assay using an Annexin V FITC/PI double stain assay (BD Biosciences) following the manufacturer's protocol.

Lin X.L., Yang L., Fu S.W., Lin W.F., Gao Y.J., Chen H.Y, & Ge Z.Z. (2017). Overexpression of NOX4 predicts poor prognosis and promotes tumor progression in human colorectal cancer. Oncotarget, 8(20), 33586-33600.

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