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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 Number | Excitation/Emission | Flow Cytometer Laser Lines | Conjugate |
|---|---|---|---|
| A13199 | 494/518 | 488 | FITC |
| A13201 | 495/519 | 488 | Alexa Fluor 488 |
| A13202 | 578/603 | 532, 561 | Alexa Fluor 568 |
| A13203 | 590/617 | 532 | Alexa Fluor 594 |
| A13204 | Biotin-X | ||
| A23202 | 346/442 | UV | Alexa Fluor 350 |
| A23204 | 650/665 | 633-637 | Alexa Fluor 647 |
| A35108 | 555/565 | 532, 561 | Alexa Fluor 555 |
| A35109 | 679/702 | 633-637 | Alexa Fluor 680 |
| A35110 | 650/660 | 633-637 | APC (Allophycocyanin) |
| A35111 | 565/578 | 488, 532, 561 | PE |
| A35122 | 410/455 | 405 | Pacific Blue |
Catalog number A13199
Price (USD)
437.00
Each
-
Excitation/Emission:
494/518
Flow Cytometer Laser Lines:
488
Conjugate:
FITC
Price (USD)
437.00
Each
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.
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
ColorGreen
DescriptionAnnexin V, FITC conjugate (replaces AnnexinV01, AnnexinV013, PHN1010, PHN1008)
Excitation/Emission494/518
Flow Cytometer Laser Lines488
For Use With (Equipment)Flow Cytometer
Kit ContentsContains 1 vial of annexin V, FITC conjugate.
No. of Reactions100
Product TypeAnnexin V conjugate
Quantity500 μL
Shipping ConditionWet Ice
ConjugateFITC
Unit SizeEach
Contents & Storage
Store in refrigerator (2°C to 8°C) and protect from light.
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Frequently asked questions (FAQs)
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.
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.
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.
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.
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.
Citations & References (449)
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Citations & References
Abstract
The p42/p44 MAP kinase pathway prevents apoptosis induced by anchorage and serum removal.
Journal:Molecular biology of the cell
PubMed ID:10712523
Anchorage removal like growth factor removal induces apoptosis. In the present study we have characterized signaling pathways that can prevent this cell death using a highly growth factorâ and anchorage-dependent line of lung fibroblasts (CCL39). After anchorage removal from exponentially growing cells, annexin V-FITC labeling can be detected after 8
Journal:
PubMed ID:18258751
Journal:
PubMed ID:10891486
Strategies for phenotyping apoptotic peripheral human lymphocytes comparing ISNT, annexin-V and 7-AAD cytofluorometric staining methods.
Journal:Journal of immunological methods
PubMed ID:9461328
The present article compares the reliability of four previously described cytofluorometric methods of apoptosis quantification for phenotyping apoptotic human lymphocytes. Each of these assays detects distinct cellular alterations of the apoptotic process. Alteration in plasma membrane integrity can be evaluated following 7-AAD incorporation and the translocation of phosphatidylserine from the
Analysis of ethanol effects on corneal epithelium.
Journal:Invest Ophthalmol Vis Sci
PubMed ID:23674759
Ethanol is widely used in ocular surface surgeries and for the treatment of corneal diseases. However, ethanol is a toxic agent that is related to the development of a number of alcohol-related diseases. Despite the common use of ethanol for therapeutic purposes in ophthalmology, effects of ethanol on the ocular
449 total citations