Annexin V-Fluorescein Apoptosis Assay Kit
Equine Annexin V-Fluorescein Apoptosis Assay Kit
Applications
Flow Cytometry, Functional Assay
Synonyms
Annexin V-Fluorescein Apoptosis; Equine Annexin V-Fluorescein Apoptosis Assay Kit; Apoptosis; Annexin V-Fluorescein Apoptosis assay kit
Applicable Applications for Annexin V-Fluorescein Apoptosis assay kit
Flow Cytometry (FC/FACS)
Application Notes
1. Prepare samples and controls.
2. Dilute 10X Binding Buffer 1:10 WITH DiH2O.
3. Wash cells twice in ice-cold culture medium or PBS and resuspend in ice-cold 1X Binding Buffer at 5 x 105-1 x 106 cells/mL.
4. Reconstitute Equine Annexin V-Fluorescein in 1 mL Annexin Reconstitution Buffer, 1X.
5. Dilute reconstituted Annexin V-Fluorescein 1:5 in PBS.
6. Add diluted Annexin V-Fluorescein to each sample at ~1:10.
7. Dilute PI 1:10 in PBS.
8. Add diluted PI to each sample at ~1:20.
9. Incubate 15 minutes on ice and protected from light. 10. Add 400 uL 1X Binding Buffer to each sample.
11. Analyze with a flow cytometer. Annexin V-Fluorescein excites at 494 nm and emits at 521 nm. PI excites at 536 nm and emits at 617 nm.
2. Dilute 10X Binding Buffer 1:10 WITH DiH2O.
3. Wash cells twice in ice-cold culture medium or PBS and resuspend in ice-cold 1X Binding Buffer at 5 x 105-1 x 106 cells/mL.
4. Reconstitute Equine Annexin V-Fluorescein in 1 mL Annexin Reconstitution Buffer, 1X.
5. Dilute reconstituted Annexin V-Fluorescein 1:5 in PBS.
6. Add diluted Annexin V-Fluorescein to each sample at ~1:10.
7. Dilute PI 1:10 in PBS.
8. Add diluted PI to each sample at ~1:20.
9. Incubate 15 minutes on ice and protected from light. 10. Add 400 uL 1X Binding Buffer to each sample.
11. Analyze with a flow cytometer. Annexin V-Fluorescein excites at 494 nm and emits at 521 nm. PI excites at 536 nm and emits at 617 nm.
Sample
Cell culture
Excitation/Emission
Annexin V-Fluorescein-494 nm / 521 nm PI-536 nm / 617 nm
Preparation and Storage
Store at 2-8 degree C.
Related Product Information for Annexin V-Fluorescein Apoptosis assay kit
Description: Equine Annexin V-Fluorescein Apoptosis Assay Kit provides a proven method for quickly and easily distinguishing two populations of dying cells from viable cells using recombinant fluorescein-conjugated Equine Annexin V and Propidium Iodide (PI). These cells will become dually labeled with Annexin V-Fluorescein (green fluorescence) and Propidium Iodide (red fluorescence). Results can be analyzed via flow cytometer.
Background: Annexin V is a member of a calcium and phospholipid binding family of proteins with vascular anticoagulant activity. Results from in vitro experiments indicate that it may play a role in the inhibition of blood coagulation by competing for phosphatidylserine (PS) binding sites with prothrombin. In healthy cells, PS is usually kept in the inner-leaflet (the cytosolic side) of the cell membrane. When a cell undergoes apoptosis, one of the earliest detectable indicators is the loss of membrane asymmetry. No longer restricted to the cytosolic part of the membrane, PS is translocated to the outer-leaflet and becomes exposed on the surface of the cell.
Equine Annexin V-Fluorescein Apoptosis Assay Kit provides a proven method for quickly and easily distinguishing two populations of dying cells from viable cells using recombinant fluorescein-conjugated Equine Annexin V and Propidium Iodide (PI). Cells with intact cell membranes and surface-exposed PS, a prominent feature of apoptosis, will stain positive for Annexin V-Fluorescein. PI is included in the kit to identify dying, later stage apoptotic cells which have lost plasma membrane integrity. These cells will become dually labeled with Annexin V-Fluorescein and Propidium Iodide (green and red fluorescence). Live non-apoptotic cells with intact plasma membranes will exclude PI and will remain unstained by the Annexin V-Fluorescein probe, assuming no treatment or cell cycle-associated event temporarily exposes the normally internalized, negatively charged PS entity. The kit also includes a specially formulated, calcium-based binding buffer which is required for Annexin V binding to occur. Analyze results using a flow cytometer.
Background: Annexin V is a member of a calcium and phospholipid binding family of proteins with vascular anticoagulant activity. Results from in vitro experiments indicate that it may play a role in the inhibition of blood coagulation by competing for phosphatidylserine (PS) binding sites with prothrombin. In healthy cells, PS is usually kept in the inner-leaflet (the cytosolic side) of the cell membrane. When a cell undergoes apoptosis, one of the earliest detectable indicators is the loss of membrane asymmetry. No longer restricted to the cytosolic part of the membrane, PS is translocated to the outer-leaflet and becomes exposed on the surface of the cell.
Equine Annexin V-Fluorescein Apoptosis Assay Kit provides a proven method for quickly and easily distinguishing two populations of dying cells from viable cells using recombinant fluorescein-conjugated Equine Annexin V and Propidium Iodide (PI). Cells with intact cell membranes and surface-exposed PS, a prominent feature of apoptosis, will stain positive for Annexin V-Fluorescein. PI is included in the kit to identify dying, later stage apoptotic cells which have lost plasma membrane integrity. These cells will become dually labeled with Annexin V-Fluorescein and Propidium Iodide (green and red fluorescence). Live non-apoptotic cells with intact plasma membranes will exclude PI and will remain unstained by the Annexin V-Fluorescein probe, assuming no treatment or cell cycle-associated event temporarily exposes the normally internalized, negatively charged PS entity. The kit also includes a specially formulated, calcium-based binding buffer which is required for Annexin V binding to occur. Analyze results using a flow cytometer.
Product Categories/Family for Annexin V-Fluorescein Apoptosis assay kit
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Product Notes
The Annexin V-Fluorescein Apoptosis (Catalog #AAA258373) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's Annexin V-Fluorescein Apoptosis can be used in a range of immunoassay formats including, but not limited to, Flow Cytometry (FC/FACS). 1. Prepare samples and controls. 2. Dilute 10X Binding Buffer 1:10 WITH DiH2O. 3. Wash cells twice in ice-cold culture medium or PBS and resuspend in ice-cold 1X Binding Buffer at 5 x 105-1 x 106 cells/mL. 4. Reconstitute Equine Annexin V-Fluorescein in 1 mL Annexin Reconstitution Buffer, 1X. 5. Dilute reconstituted Annexin V-Fluorescein 1:5 in PBS. 6. Add diluted Annexin V-Fluorescein to each sample at ~1:10. 7. Dilute PI 1:10 in PBS. 8. Add diluted PI to each sample at ~1:20. 9. Incubate 15 minutes on ice and protected from light. 10. Add 400 uL 1X Binding Buffer to each sample. 11. Analyze with a flow cytometer. Annexin V-Fluorescein excites at 494 nm and emits at 521 nm. PI excites at 536 nm and emits at 617 nm. Researchers should empirically determine the suitability of the Annexin V-Fluorescein Apoptosis 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 "Annexin V-Fluorescein Apoptosis, Assay Kit" 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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