Annexin V Conjugates for Apoptosis Detection
Annexin V Conjugates for Apoptosis Detection
Citations & References (14)
Invitrogen™

Annexin V Conjugates for Apoptosis Detection

Detect early stages of apoptosis with Annexin V stand-alone Alexa Fluor, APC, Pacific Blue, PE, FITC, and biotin conjugates using flow cytometry.
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Catalog NumberExcitation/EmissionFlow Cytometer Laser LinesConjugate
A13203590/617532Alexa Fluor 594
A13201495/519488Alexa Fluor 488
A13199494/518488FITC
A13202578/603532, 561Alexa Fluor 568
A13204Biotin-X
A23202346/442UVAlexa Fluor 350
A23204650/665633-637Alexa Fluor 647
A35108555/565532, 561Alexa Fluor 555
A35109679/702633-637Alexa Fluor 680
A35110650/660633-637APC (Allophycocyanin)
A35111565/578488, 532, 561PE
A35122410/455405Pacific Blue
Catalog number A13203
Price (JPY)
112,600
Each
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Excitation/Emission:
590/617
Flow Cytometer Laser Lines:
532
Conjugate:
Alexa Fluor 594
Achieve quick and reliable detection of early cell apoptosis with Annexin V stand-alone conjugates for apoptosis detection. Annexin V conjugates offer up to 100-fold difference in fluorescence signal intensity between apoptotic and non-apoptotic cells using flow cytometry.
Annexin V has a high affinity for phosphatidylserine (PS), which becomes exposed on the outer leaflet of cells undergoing apoptosis. Because of this affinity, fluorescently labeled annexin V reagents are commonly used in apoptosis research.

Annexin V conjugates provide quick and reliable detection methods for studying the externalization of phosphatidylserine, an indicator of intermediate stages of apoptosis. The difference in fluorescence intensity between apoptotic and nonapoptotic cells stained with our fluorescent annexin V conjugates, as measured by flow cytometry, is typically about 100-fold.

In collaboration with Nexins Research BV, we provide the best and brightest annexin V conjugates available, including Alexa Fluor 350, 488, 555, 568, 594, 647, and 680 annexin V conjugates, as well as Annexin V APC, Biotin-X, FITC, Pacific Blue, and PE conjugates. Highly fluorescent annexin V conjugates provide quick and reliable detection methods for studying the externalization of phosphatidylserine, one of the earliest indicators of apoptosis.

The Annexin V Pacific Blue conjugate is violet excitable, making it ideal for instruments with a violet laser and for multicolor experiments that include green- or red-fluorescent dyes.

The benefits of our annexin V conjugates include:
• Conjugated to Invitrogen Alexa Fluor and eFluor dyes for brighter signals
• Conjugates for all available lasers
• Available as stand-alone reagents or easy-to-use kits

Annexin V staining to detect apoptotic cells can only be done on live cells and tissue. If samples are to be fixed post-staining, there are specific conditions required to achieve transient retention of signal. These include use of an alcohol-free, aldehyde-based fixation method, use of buffers containing Ca2+ and avoidance of surfactants/detergents. For your convenience, we also offer a concentrated annexin-binding buffer that facilitates the binding of annexin V to phosphatidylserine in apoptosis assays.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
ColorRed
DescriptionAnnexin V, Alexa Fluor 594 conjugate
Excitation/Emission590/617
Flow Cytometer Laser Lines532
For Use With (Equipment)Flow Cytometer
Kit ContentsContains 1 vial of annexin V, Alexa Fluor 594 conjugate.
No. of Reactions100
Product TypeAnnexin V conjugate
Quantity500 μL
Shipping ConditionWet Ice
ConjugateAlexa Fluor 594
Unit SizeEach
Contents & Storage
Store in refrigerator (2°C to 8°C) and protect from light.

Frequently asked questions (FAQs)

I want to study apoptosis using an Annexin V conjugate, but with adherent cells via microscopy instead of flow cytometry. Can this be done?

It has been done, but we don‘t recommend it. Both healthy cells and apoptotic cells possess phosphatidylserine on the cell surface, which can be detected with Annexin V, but apoptotic cells have significantly more of it. You can easily tell the difference between these two populations with flow cytometry, because flow cytometers are more sensitive and have a higher throughput. But with a microscope, you cannot always tell the difference, especially for adherent cells. Instead, for microscopy, we recommend a different technique, such as detecting caspases with CellEvent Caspase Detection Reagents.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I trypsinized my adherent cells and labeled with annexin V, and now my flow data is showing a high percentage of apoptotic cells even for control, untreated cells. What is the problem?

Trypsinization or mechanical scraping of cells temporarily disrupts the plasma membrane, allowing annexin V to bind phosphatidylserine on the cytoplasmic surface of the cell membrane and thus leading to false positive staining. Allow the cells to recover for about 30 minutes in optimal cell culture conditions and medium after trypsinizing/scraping so that they can recover their membrane integrity before staining. For lightly adherent cell lines, such as HeLa and NIH 3T3, another option is to use non-enzyme treatments like Gibco Cell Dissociation Buffer (Cat. No. 13151014).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I detect annexin V staining in an imaging assay?

Annexin V staining is not typically used in imaging experiments; it is a better reagent for flow cytometry analysis. All cells will stain to some extent, so it can be difficult to distinguish a relatively bright annexin V-stained cell from a dimmer non-apoptotic cell. Caspase activation, detected using our CellEvent Caspase 3/7 or Image-iT LIVE Caspase detection kits, is a better method for detecting apoptosis in an imaging assay.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

When should I stain adherent cells with annexin V for flow cytometric analysis? Before or after I trypsinize them?

Trypsinize first and then allow the cells to recover about 30 minutes in optimal cell culture conditions and medium before staining with annexin V conjugates. Trypsinization or mechanical scraping of cells temporarily disrupts the plasma membrane, allowing for annexin V to bind phosphatidylserine on the cytoplasmic surface of the cell membrane and thus leading to false positive staining. For lightly adherent cell lines such as HeLa and NIH 3T3, you could use a less harsh (non-enzymatic) dissociation product like Gibco Cell Dissociation Buffer (Cat. No. 13151014).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I fix my cells after annexin V labeling?

Annexin V staining is best analyzed on live cells. If you need to fix your cells for analysis, then fix in 3.7% formaldehyde in PBS containing calcium and magnesium to maintain binding during fixation. The signal will not be retained after permeabilization, thus annexin V staining is not compatible with internal antibody labeling.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

View all Annexin V Conjugates for Apoptosis Detection FAQs

Citations & References (14)

Citations & References
Abstract
Caspase-mediated cleavage of the stacking protein GRASP65 is required for Golgi fragmentation during apoptosis.
Authors:Lane JD, Lucocq J, Pryde J, Barr FA, Woodman PG, Allan VJ, Lowe M
Journal:J Cell Biol
PubMed ID:11815631
'The mammalian Golgi complex is comprised of a ribbon of stacked cisternal membranes often located in the pericentriolar region of the cell. Here, we report that during apoptosis the Golgi ribbon is fragmented into dispersed clusters of tubulo-vesicular membranes. We have found that fragmentation is caspase dependent and identified GRASP65 ... More
A novel role for microtubules in apoptotic chromatin dynamics and cellular fragmentation.
Authors:Moss DK, Betin VM, Malesinski SD, Lane JD
Journal:J Cell Sci
PubMed ID:16723742
'Dramatic changes in cellular dynamics characterise the apoptotic execution phase, culminating in fragmentation into membrane-bound apoptotic bodies. Previous evidence suggests that actin-myosin plays a dominant role in apoptotic cellular remodelling, whereas all other cytoskeletal elements dismantle. We have used fixed cells and live-cell imaging to confirm that interphase microtubules rapidly ... More
Distinct phosphoinositide 3-kinases mediate mast cell degranulation in response to G-protein-coupled versus FcepsilonRI receptors.
Authors:Windmiller DA, Backer JM
Journal:J Biol Chem
PubMed ID:12529321
'Phosphoinositide (PI) 3-kinases are critical regulators of mast cell degranulation. The Class IA PI 3-kinases p85/p110beta and p85/p110delta but not p85/p110alpha are required for antigen-mediated calcium flux in RBL-2H3 cells (Smith, A. J., Surviladze, Z., Gaudet, E. A., Backer, J. M., Mitchell, C. A., and Wilson, B. S. et al., ... More
Cholesterol-induced apoptotic macrophages elicit an inflammatory response in phagocytes, which is partially attenuated by the Mer receptor.
Authors:Li Y, Gerbod-Giannone MC, Seitz H, Cui D, Thorp E, Tall AR, Matsushima GK, Tabas I
Journal:J Biol Chem
PubMed ID:16380374
'Macrophage apoptosis and the ability of phagocytes to clear these apoptotic cells are important processes in advanced atherosclerosis. Phagocytic clearance not only disposes of dead cells but usually elicits an anti-inflammatory response. To study this process in a model of advanced lesional macrophage death, macrophages rendered apoptotic by free cholesterol ... More
CD45-mediated fodrin cleavage during galectin-1 T cell death promotes phagocytic clearance of dying cells.
Authors:Pang M, He J, Johnson P, Baum LG,
Journal:J Immunol
PubMed ID:19454697
'Disassembly and phagocytic removal of dying cells is critical to maintain immune homeostasis. The factors that regulate fragmentation and uptake of dying lymphocytes are not well understood. Degradation of fodrin, a cytoskeletal linker molecule that attaches CD45 to the actin cytoskeleton, has been described in apoptotic cells, although no specific ... More
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