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Référence | Excitation / émission | Faisceaux laser du cytomètre en flux | Conjugué |
---|---|---|---|
A35111 | 565/578 | 488, 532, 561 | PE |
A13201 | 495/519 | 488 | Alexa Fluor 488 |
A13199 | 494/518 | 488 | FITC |
A13202 | 578/603 | 532, 561 | Alexa Fluor 568 |
A13203 | 590/617 | 532 | Alexa Fluor 594 |
A13204 | Biotine-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 (Allophycocyanine) |
A35122 | 410/455 | 405 | Pacific Blue |
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.
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.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
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