eBioscience™ Annexin V Apoptosis Detection Kits
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Invitrogen™

eBioscience™ Annexin V Apoptosis Detection Kits

Kits utilize viability dye-conjugated annexin V to label phosphatidylserine (PS), an early marker of apoptosis, on the extracellular membrane.
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Catalog NumberConjugateExcitation/EmissionQuantity
BMS500BT-300
also known as BMS500BT/300
Biotin, Propidium IodideNA, 535/NA, 617300 Tests
88-8005-74FITC, Propidium Iodide488, 535/520, 617200 Tests
88-8006-72eFluor 450, 7-AAD405, 546/450, 64750 Tests
88-8006-74eFluor 450, 7-AAD405, 546/450, 647200 Tests
88-8007-74APC, Propidium Iodide650, 535/660, 617200 Tests
88-8008-72PCP-eFluor 71050 Tests
88-8008-74PCP-eFluor 710200 Test
BMS500FI-100FITC, Propidium Iodide488, 535/520, 617100 Tests
BMS500FI-20FITC, Propidium Iodide488, 535/520, 61720 Tests
BMS500FI-300FITC, Propidium Iodide488, 535/520, 617300 Tests
88-8102-72PE, 7-AAD50 Tests
88-8102-74PE, 7-AAD200 Tests
88-8103-72PE/Cy750 Tests
88-8103-74PE/Cy7200 Tests
BMS500BT-100
also known as BMS500BT/100
Biotin, Propidium IodideNA, 535/NA, 617100 Tests
BMS500BT-20
also known as BMS500BT/20
Biotin, Propidium IodideNA, 535/NA, 61720 Tests
88-8005-72FITC, Propidium Iodide488, 535/520, 61750 Tests
88-8007-72APC, Propidium Iodide650, 535/660, 61750 Tests
Catalog number BMS500BT-300
also known as BMS500BT/300
Price (USD)
564.00
Each
Add to cart
Conjugate:
Biotin, Propidium Iodide
Excitation/Emission:
NA, 535/NA, 617
Quantity:
300 Tests
Price (USD)
564.00
Each
Add to cart
Easily distinguish cells undergoing early and late apoptosis with eBioscience and Annexin V Apoptosis Detection kits for flow cytometry analysis. These kits utilize viability dye-conjugated annexin V to label phosphatidylserine (PS), an early marker of apoptosis, on the extracellular membrane. In late-stage apoptosis, as cell integrity is lost, both annexin V and the viability dye traverse and bind PS on the interior of the plasma membrane.
Annexins are a family of calcium-dependent phospholipid-binding proteins that preferentially bind phosphatidylserine (PS). Under normal physiologic conditions, PS is predominantly located in the inner leaflet of the plasma membrane. Upon initiation of apoptosis, PS loses its asymmetric distribution across the phospholipid bilayer and is translocated to the extracellular membrane leaflet marking cells as targets of phagocytosis. Once on the outer surface of the membrane, PS can be detected by fluorescently labeled annexin V in a calcium-dependent manner.

In early-stage apoptosis, the plasma membrane excludes viability dyes such as propidium iodide (PI), 7-AAD, or fixable viability dyes such as eFluor 450. These cells will stain with annexin V but not a viability dye, thus distinguishing cells in early apoptosis. However, in late-stage apoptosis, the cell membrane loses integrity, thereby allowing annexin V to also access PS in the interior of the cell. A viability dye can be used to resolve these late-stage apoptotic and necrotic cells (annexin V, viability dye-positive) from the early-stage apoptotic cells (annexin V positive, viability dye-negative).

Current viability stains available with the eBioscience Annexin V Apoptosis Detection kits include FITC, eFluor 450, allophycocyanin (APC), phycoerythrin (PE), PE-cyanine7, and biotin.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
DescriptioneBioscience Annexin V-Biotin Apoptosis Detection Kit
Quantity300 Tests
Kit ContentsContains:
• Binding Buffer (4X), 2 bottles (50 mL)
• rh Annexin V-Biotin, 1 vial (1.5 mL)
• Propidium Iodide (20 μg/mL), 2 vials (1.8 mL)
Product TypeApoptosis Detection Kit
Excitation/EmissionNA, 535/NA, 617
Shipping ConditionWet Ice
ConjugateBiotin, Propidium Iodide
For Use With (Equipment)Flow Cytometer
Unit SizeEach
Contents & Storage
Handling Conditions:
Store at 2-8°C. Avoid temperature fluctuations.

Frequently asked questions (FAQs)

Is annexin V specific for any particular species?

Annexin V binding of phospholipids is not species-specific. In the presence of millimolar concentrations of Ca2+, Annexin V binds to several phospholipids, but exhibits the highest affinity for phosphatidylserine. This feature is useful for screening cells undergoing apoptosis. Phosphatidylserine is usually distributed on the inner leaflet of the lipid bilayer, but when cells start to undergo apoptosis, the phosphatidylserine moves to the outer leaflet. The translocation of phosphatidylserine to the outer leaflet makes it available for Annexin V staining. Normal, intact cells will not stain with Annexin V, while cells undergoing apoptosis will be.

Annexin V staining can be visualized by fluorescence microscopy or flow cytometry using conjugates such as Annexin V-FITC, or Annexin V-biotin in conjugation with a streptavidin-conjugated fluorophore. It is important to note that experiments with Annexin V require that the cells under study are live cells with intact membranes. It is not suitable for use with fixed cells as the membrane develops holes during the fixation process, making the phosphatidylserine on the inner face of the membrane accessible for Annexin V binding. For similar reasons, this product is also not suitable for use with tissue sections. Because necrotic cells also have numerous membrane breaches, these cells are also often stained with Annexin V. A quick way to differentiate apoptotic from necrotic cells is with a double staining regime using Annexin V-FITC (green fluorescence) and the DNA staining dye propidium iodide (red fluorescence). Propidium iodide requires membrane breaches in order to gain access to the cell nucleus.

Using the Annexin V-FITC/propidium iodide double staining regime, the following staining patterns are observed with apoptotic and necrotic cells:
1. Cells that are normal will not stain with either Annexin V-FITC or propidium iodide (indicating no phosphatidylserine on the surface and no holes in the membrane).
2. Cells that are starting to undergo apoptosis will stain with Annnexin V-FITC only (indicating phosphatidylserine on the surface but no holes in the membrane).
3. Cells that are in late apoptosis and necrotic cells will stain with both Annexin V-FITC and with propidium iodide (indicating that we cannot tell where the phosphatidylserine is located and that there are holes in the membrane).

Phosphatidylserine is an ubiquitous membrane component among mammalian species; therefore, we believe that Annexin V has utility in monitoring apoptosis with any mammalian-derived cell line. One report (O'Brien et al., 1997) has even indicated its utility in monitoring apoptosis with plant cells. Dive, C., C.D. Gregory, D.J. Phipps, D.L. Evans, A.E. Milner, and A.H. Wyllie (1992) Analysis and discrimination of necrosis and apoptosis (programmed cell death) by multiparameter flow cytometry. Biochem. Biophys. Acta 1133:275-285. Koopman, G., C.P.M. Reutlingsperger, G.A.M. Kuitjen, R.M.J. Keehnen, S.T. Pals, and M.H.J. van Oers (1994) Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84:1415-1420. O'Brien, I.E., C.P. Reutelingsperger, and K.M. Holdaway (1997) Annexin-V and TUNEL use in monitoring the progression of apoptosis in plants. Cytometry 29(1):28-33.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Can I use eFluor 450-conjugated antibodies with eBioscience Annexin V Apoptosis Detection kits?

Yes, eFluor 450-conjugated antibodies are compatible with eBioscience Annexin V Apoptosis Detection kits.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.