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Molecular Probes™ Annexin V Conjugates for Apoptosis Detection
Description
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 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.
Specifications
Specifications
| Description | Annexin V, Alexa Fluor 680 conjugate |
| Quantity | 500 μL |
| Product Type | Annexin V conjugate |
| Content And Storage | Contains 1 vial of annexin V, Alexa Fluor 680 conjugate. Store in refrigerator (2°C to 8°C) and protect from light. |
| Flow Cytometer Laser Lines | 633-637 |
| Excitation/Emission | 679/702 |
| No. of Reactions | 100 |
| Color | Near IR |
| Shipping Condition | Wet Ice |
| Conjugate | Alexa Fluor 680 |
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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.
Treating cells with trypsin or other reagents to detach adherent cells causes damage to the membrane, such that cells will be labeled with annexin V. The best way to avoid this problem is to allow your cells to recover for 30-45 min in the incubator. Swirl the tube/plate/flask every few minutes to prevent re-attachment. After this recovery period, you can label your cells with annexin V and analyze by flow cytometry.
-Measures data from single cells.
-Data are obtained for a large number of cells, generating a rich statistical analysis of cell populations.
-Because single cells are measured, it will reveal heterogeneity within a population.
-With the ability to multiplex, small sub-populations can be identified.
-Thousands of cells can be analyzed rapidly.
-It is ideally suited for blood samples and other cells in suspension.
-Data can be re-analyzed multiple times after acquisition.
-Flow cytometry files (FCS) can be archived.
There are several applications, some of which include immunophenotyping, cell cycle analysis, apoptosis assays such as annexin V staining, CellEvent Caspase-3/7 assay, and TUNEL assay, cell viability, proliferation assays such as CellTrace assay and Click-iT EdU assay, measurements of mitochondrial potential with MitoProbe assays, and cell counting using counting beads.
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