Annexin V | AAT Bioquest

Annexin V-iFluor® 647 conjugate

Annexin V-iFluor® 647 Conjugate is a far-red fluorescent probe for detecting phosphatidylserine exposure on apoptotic cells using flow cytometry and fluorescence microscopy.

Fluorescence Detection: Excitation at 656 nm and emission at 670 nm.
Apoptosis Detection: Detects phosphatidylserine exposure during early-stage apoptosis.
Calcium-Dependent Binding: Annexin V binds exposed phosphatidylserine in the presence of calcium.
Instrument Compatibility: Supports 640 nm laser and Cy5 filter set.
Multiplex Compatibility: Compatible with dead-cell stains for distinguishing apoptotic and membrane-compromised cells.

The detection of binding activity of Annexin V-iFluor® 647 and phosphatidylserine in Jurkat cells. Jurkat cells were treated without (Blue) or with 20 µM staurosporine(Red) in a 37 °C, 5% CO2 incubator for 4-5 hours, and then dye loaded with Annexin V-iFluor® 647 for 30 minutes. The fluorescence intensity of Annexin V-iFluor® 647 was measured with a FACSCalibur (Becton Dickinson) flow cytometer using the FL4 channel.

Protocol

SDS

Catalog	Size	Price	Quantity
20074	100 tests	Price

Overview

Annexins are a family of proteins that bind to phospholipid membranes in the presence of calcium. Annexin V is a valuable tool for studying cell apoptosis. It is used as a probe to detect cells which have expressed phosphatidylserine on the cell surface, a feature found in apoptosis as well as other forms of cell death. There are a variety of parameters that can be used for monitoring cell viability. Annexin V-dye conjugates are widely used to monitor cell apoptosis through measuring the translocation of phosphatidylserine (PS). In apoptosis, PS is transferred to the outer leaflet of the plasma membrane. The appearance of phosphatidylserine on the cell surface is a universal indicator of the initial/intermediate stages of cell apoptosis and can be detected before morphological changes can be observed. This fluorescent Annexin V conjugate has spectral properties similar to Alexa Fluor® 647 (Alexa Fluor® 647 is the trademark of Invitrogen) and Cy5® (Cy5® is the trademark of GE Healthcare).

Citations

View all 7 citations: Citation Explorer

The mitochondrial NADH shuttle system is a targetable vulnerability for Group 3 medulloblastoma in a hypoxic microenvironment

Authors:

Contenti, J and Guo, Y and Mazzu, A and Irondelle, M and Rouleau, M and Lago, C and Leva, G and Tiberi, L and Ben-Sahra, I and Bost, F and others,

Journal:

Cell Death \& Disease (2023): 1--12

An anti-mesothelin targeting antibody drug conjugate induces pyroptosis and ignites antitumor immunity in mouse models of cancer

Authors:

Wittwer, Nicole L and Staudacher, Alexander H and Liapis, Vasilios and Cardarelli, Pina and Warren, Harriet and Brown, Michael P

Journal:

Journal for ImmunoTherapy of Cancer (2023): e006274

Microvesicles in human reproduction and their role in Assisted Reproductive Techniques

Authors:

Sawas, Hala

Glial signaling mechanisms in the progression of neuroinflammatory injury

Authors:

Popichak, Katriana A

Glial-neuronal signaling mechanisms underlying the neuroinflammatory effects of manganese

Authors:

Popichak, Katriana A and Afzali, Maryam F and Kirkley, Kelly S and Tjalkens, Ronald B

Journal:

Journal of neuroinflammation (2018): 324

References

View all 32 references: Citation Explorer

Gold fluorescent annexin A5 as a novel apoptosis detection tool

Authors:

Kurschus FC, Pal PP, Baumler P, Jenne DE, Wiltschi B, Budisa N.

Journal:

Cytometry A (2009): 626

Glycogen synthase kinase-3 and Omi/HtrA2 induce annexin A2 cleavage followed by cell cycle inhibition and apoptosis

Authors:

Wang CY, Lin YS, Su WC, Chen CL, Lin CF.

Journal:

Mol Biol Cell (2009): 4153

Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection

Authors:

Palma PF, Baggio GL, Spada C, Silva RD, Ferreira SI, Treitinger A.

Journal:

Braz J Infect Dis (2008): 108

Measurement of annexin V uptake and lactadherin labeling for the quantification of apoptosis in adherent Tca8113 and ACC-2 cells

Authors:

Hu T, Shi J, Jiao X, Zhou J, Yin X.

Journal:

Braz J Med Biol Res (2008): 750

Detection of apoptosis induced by new type gosling viral enteritis virus in vitro through fluorescein annexin V-FITC/PI double labeling

Authors:

Chen S, Cheng AC, Wang MS, Peng X.

Journal:

World J Gastroenterol (2008): 2174

Physical properties

Molecular weight	~36000
Solvent	Water

Spectral properties

Correction factor (260 nm)	0.03
Correction factor (280 nm)	0.03
Correction factor (656 nm)	0.0793
Extinction coefficient (cm ^-1 M ^-1)	220000 ¹
Excitation (nm)	655
Emission (nm)	670
Quantum yield	0.25 ¹

Usage and storage

Intended use	Research Use Only (RUO)
Storage	Freeze (< -15 °C); Minimize light exposure

Instrument settings

Flow cytometer
Excitation	640 nm laser
Emission	660/20 nm filter
Instrument specification(s)	APC channel

Fluorescence microscope
Excitation	Cy5 filter set
Emission	Cy5 filter set
Recommended plate	Black wall/clear bottom

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Page updated on October 2, 2026