Annexin V Conjugates for Apoptosis Detection
Use with Annexin Binding Buffer for flow cytometry
Annexin V Conjugates for Apoptosis Detection
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 NumberConjugateExcitation/EmissionFlow Cytometer Laser Lines
A35122Pacific Blue410/455405
A13201Alexa Fluor 488495/519488
A13199FITC494/518488
A13202Alexa Fluor 568578/603532, 561
A13203Alexa Fluor 594590/617532
A13204Biotin-X
A23202Alexa Fluor 350346/442UV
A23204Alexa Fluor 647650/665633-637
A35108Alexa Fluor 555555/565532, 561
A35109Alexa Fluor 680679/702633-637
A35110APC (Allophycocyanin)650/660633-637
A35111PE565/578488, 532, 561
Catalog number A35122
Price (USD)
448.00
Each
Add to cart
Conjugate:
Pacific Blue
Excitation/Emission:
410/455
Flow Cytometer Laser Lines:
405
Price (USD)
448.00
Each
Add to cart
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
DescriptionAnnexin V, Pacific Blue conjugate, for flow cytometry
Kit ContentsContains 1 vial of annexin V, Pacific Blue conjugate.
Product TypeAnnexin V conjugate
Flow Cytometer Laser Lines405
Excitation/Emission410/455
No. of Reactions100
ColorBlue
Shipping ConditionWet Ice
ConjugatePacific Blue
For Use With (Equipment)Flow Cytometer
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.

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