BioReagent, Biological Stain, for microscopy, Suitable for Immunofluorescence(IF), 5 μL/test Biological Stain,BioReagent,für die Mikroskopie,Suitable for Immunofluorescence(IF) for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
Store at 2-8°C,Protected from light,Do not freeze Ships Wet ice,Do not freeze Check lot-specific COA for exact specifications.
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Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
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. 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).
We offer recombinant Annexin V conjugated to a numerous fluorophores, as well as an Annexin V biotin conjugate which can be detected with fluorophore-labeled streptavidin. By binding to PS, fluorophores labeled Annexin V can be used to detect and quantify apoptotic cells via flow cytometry or fluorescence microscopy. The excitation and emission maxima of the Annexin V conjugates are summarized in the following table.
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Precautions
1. Please try to avoid light when using to slow down the quenching of fluorescence.
2. Propidium Iodide Solution is toxigenic and mutagenic; handle with care.
3. Due to the calcium dependence of the Annexin V:PS interaction, it is critical to avoid buffers containing EDTA or other calcium chelators during Annexin V experiments.
Instruction for use
1. Dilute 10X Binding Buffer (A1372288) to 1X using distilled water (1 mL 10X Binding Buffer + 9 mL ddH2O).
2. Wash cells twice with cold PBS and then resuspend the desired amounts of cells in Annexin V Binding Buffer at a concentration of 1.0-5.0 x 106 cells/mL.
3. Add 5 µL of Annexin V-AF594 to 100 µL of the cell suspension. Stain with a viability dye, such as PI (P1373641; P1372285), 7-AAD (A1372406), or DAPI (D1372407) dyes, if desired.
4. Gently vortex the cells and incubate for 10 min at RT (25°C) in the dark.
5. Add 100 µL of 1X Binding Buffer to each assay. Analyze by flow cytometry within 1 hr.
Annexin V-AF594 (rp226001) - Flow Cytometry
U937 cells were either untreated (left) or treated (right) with 5µM camptothecin (C111281) for 4 hours, then stained with Annexin V-AF594 (rp226001) in Annexin V Binding buffer (A1372288) for 10 minutes at room temperature.
Annexin V-AF594 (rp226001) - Flow Cytometry
U937 cells were either untreated (blue) or treated (red) with 5µM camptothecin (C111281) for 4 hours, then stained with Annexin V-AF594 (rp226001) in Annexin V Binding buffer (A1372288) for 10 minutes at room temperature.
| Anwendung | Dilution info |
|---|---|
| Flow Cytometry | 5 μL/test; For flow cytometric staining, the suggested use of this reagent is 5 µL per 100,000 - million cells in a 100 µL volume of Annexin V Binding Buffer. It |
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