Testing Data
Aposcreen Annexin V Antibody
Aposcreen Annexin V-FITC
Applications
Flow Cytometry
Synonyms
Aposcreen Annexin V; Aposcreen Annexin V-FITC; ApoScreen Annexin V; anti-Aposcreen Annexin V antibody
Form/Format
Annexin V Fluorescein (FITC) Conjugate
Applicable Applications for anti-Aposcreen Annexin V antibody
Blocking Annexin V-conjugates, Flow Cytometry, Fluorescence Microscopy
Application Notes
Suggested Staining Protocol: 1. Wash cells twice in cold PBS; remove the PBS from the cell pellet after the second wash.
2. Resuspend cells in cold binding buffer (e.g. 10 mM HEPES pH 7.4, 140 mM NaCl, 2.5 mM CaCl2, 0.1% BSA) to a concentration of 1 x 106 to 1 x 107 cells/mL.
3. Add 100 uL of cells (1 x 105 to 1 x 106) to each of four labeled tubes.
4. Add 10 uL of labeled annexin V to tube #2 and tube #4.
5. Gently vortex each tube and incubate for 15 minutes on ice, protected from light.
6. Without washing, add 380 uL of cold 1X binding buffer to each tube. Add 10 uL of PI (from a 50 ug/ml stock solution) to tube #3 and tube #4.
7. Immediately analyze by flow cytometry. Note: the 4-tube protocol above is for initially adjusting settings on the flow cytometer; subsequent assays should basically require the protocol described for "tube #4," i.e. cells stained with both labeled annexin V and PI.
Flow Cytometry: 10 ul/10^6 cells
2. Resuspend cells in cold binding buffer (e.g. 10 mM HEPES pH 7.4, 140 mM NaCl, 2.5 mM CaCl2, 0.1% BSA) to a concentration of 1 x 106 to 1 x 107 cells/mL.
3. Add 100 uL of cells (1 x 105 to 1 x 106) to each of four labeled tubes.
4. Add 10 uL of labeled annexin V to tube #2 and tube #4.
5. Gently vortex each tube and incubate for 15 minutes on ice, protected from light.
6. Without washing, add 380 uL of cold 1X binding buffer to each tube. Add 10 uL of PI (from a 50 ug/ml stock solution) to tube #3 and tube #4.
7. Immediately analyze by flow cytometry. Note: the 4-tube protocol above is for initially adjusting settings on the flow cytometer; subsequent assays should basically require the protocol described for "tube #4," i.e. cells stained with both labeled annexin V and PI.
Flow Cytometry: 10 ul/10^6 cells
Characterization
To insure acceptable performance, each batch of product is tested by flow cytometry to conform to the characteristics of a standard reference reagent.
Warning
Reagents contain sodium azide which is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Preparation and Storage
Annexin V-FITC: Store at 2-8 degree C; protect from light
Testing Data
Testing Data
Related Product Information for anti-Aposcreen Annexin V antibody
A number of changes in cell structure occur during apoptosis. One major change is the loss of membrane phospholipid asymmetry, with translocation of phosphatidylserine (PS) from the inner leaflet of the phospholipid bilayer to the external surface.1 Annexin V (Mr 36-kDa), a member of the annexin family of calcium-dependent phospholipid binding proteins, has a high affinity for PS-containing phospholipid bilayers.2,3 When PS is exposed on the extracellular face of a cell membrane, annexin V binds with an affinity of approximately 7 nM.4 Fluorochrome conjugates of annexin V can be used to monitor changes in cell membrane phospholipid asymmetry,5-8 thereby providing a convenient tool for detection of apoptotic cells. ApoScreen Annexin V is a labeled annexin V that can be used in conjunction with propidium iodide to evaluate subpopulations of cells along the apoptotic pathway. ApoScreen detection reagents can also be used to analyze changes in membrane asymmetry in platelets5 and erythrocytes.8 Analysis can be performed on a flow cytometer equipped with a single laser using excitation at 488 nm. Alternatively, the cells can be examined with a fluorescence microscope equipped with appropriate filters.
References
1. Fadok, V.A. et al. 1992. J. Immunol. 148:2207
2. Tait, J.F., et al. 1989. J. Biol. Chem. 264:7944.
3. Andree, H.A., et al. 1990. J. Biol. Chem. 265:4923.
4. Thiagarajan, P., et al. 1990. J. Biol. Chem. 265:17420.
5. Dachary, P.J., et al. 1993. Blood 81:2554.
6. Koopman, G., et al. 1994. Blood 84:1415.
7. Martin, S.J., et al. 1995. J. Exp. Med. 182:1545.
8. Wood, B.L., et al. 1996. Blood 88:1873.
2. Tait, J.F., et al. 1989. J. Biol. Chem. 264:7944.
3. Andree, H.A., et al. 1990. J. Biol. Chem. 265:4923.
4. Thiagarajan, P., et al. 1990. J. Biol. Chem. 265:17420.
5. Dachary, P.J., et al. 1993. Blood 81:2554.
6. Koopman, G., et al. 1994. Blood 84:1415.
7. Martin, S.J., et al. 1995. J. Exp. Med. 182:1545.
8. Wood, B.L., et al. 1996. Blood 88:1873.
Similar Products
Product Notes
The Aposcreen Annexin V (Catalog #AAA670379) is an Antibody and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's Aposcreen Annexin V can be used in a range of immunoassay formats including, but not limited to, Blocking Annexin V-conjugates, Flow Cytometry, Fluorescence Microscopy. Suggested Staining Protocol: 1. Wash cells twice in cold PBS; remove the PBS from the cell pellet after the second wash. 2. Resuspend cells in cold binding buffer (e.g. 10 mM HEPES pH 7.4, 140 mM NaCl, 2.5 mM CaCl2, 0.1% BSA) to a concentration of 1 x 106 to 1 x 107 cells/mL. 3. Add 100 uL of cells (1 x 105 to 1 x 106) to each of four labeled tubes. 4. Add 10 uL of labeled annexin V to tube #2 and tube #4. 5. Gently vortex each tube and incubate for 15 minutes on ice, protected from light. 6. Without washing, add 380 uL of cold 1X binding buffer to each tube. Add 10 uL of PI (from a 50 ug/ml stock solution) to tube #3 and tube #4. 7. Immediately analyze by flow cytometry. Note: the 4-tube protocol above is for initially adjusting settings on the flow cytometer; subsequent assays should basically require the protocol described for "tube #4," i.e. cells stained with both labeled annexin V and PI. Flow Cytometry: 10 ul/10^6 cells. Researchers should empirically determine the suitability of the Aposcreen Annexin V for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Aposcreen Annexin V, Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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