Annexin Binding Buffer (5X), for flow cytometry
Annexin Binding Buffer (5X), for flow cytometry
Invitrogen™

Annexin Binding Buffer (5X), for flow cytometry

This product is designed to facilitate the binding of annexin V to phosphatidylserine for use in apoptosis assays.Read more
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Catalog NumberQuantityPackaging
V1324650 mL
Catalog number V13246
Price (USD)
91.50
Each
Add to cart
Quantity:
50 mL
Price (USD)
91.50
Each
Add to cart
This product is designed to facilitate the binding of annexin V to phosphatidylserine for use in apoptosis assays.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Quantity50 mL
pH7.4
Product TypeAnnexin Binding Buffer
Shipping ConditionWet Ice
Formulation50 mM HEPES, 700 mM NaCl, 12.5 mM CaCl2, pH 7.4
For Use With (Application)Flow Cytometry
FormConcentrated
Unit SizeEach
Contents & Storage
Contains 1 bottle of annexin binding buffer (5x solution, 50 mL).

Store at 2–8°C.

Frequently asked questions (FAQs)

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.