Annexin V-FITC/PI Apoptosis Detection Kit - Suitable for Immunofluorescence(IF), BioReagent, Biological Stain, for microscopy, high purity

Cat. No.: A1372286

GRADE & PURITY BioReagent, Biological Stain, for Microscopy, Suitable for Immunofluorescence(IF)

Overview

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).

This kit is suitable for the identification and enumeration of dead cells, such as apoptotic or necrotic cells, by flow cytometry.

A1372286	Components	20T	50T	100T	Storage	Quantity Per Test
A1372286A	10x Annexin V Binding Buffer	5mL	10mL	20mL	2-8℃	200 μL/0.5-1.0x10⁵ cells
A1372286B	Annexin V-FITC	100 μL	250 μL	500 μL	2-8℃. Store in the dark.	5 μL/0.5-1.0x10⁵ cells
A1372286C	PI Staining Solution	100 μL	250 μL	500 μL	2-8℃. Store in the dark.	2 μL/0.5-1.0x10⁵ cells

Note: The recommended number of cells to stain per test is 0.5-1.0x10⁵ cells.

Instruction for use

1. Dilute 10x Binding Buffer 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 amount of cells in Annexin V Binding Buffer at a concentration of 0.5-1.0 x 106 cells/mL.

3. Add 5 µl of Annexin-FITC and 2 µl PI to 100 µL of the cell suspension

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.

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.

Storage and Shipping

Storage

Store at 2-8°C,Protected from light,Do not freeze

Shipped In

Wet ice,Do not freeze

Stability And Storage

Store at 2-8°C long term (12 months). Store in the dark. Do not freeze.

Contents & Storage

A1372286	Components	20T	50T	100T	Storage	Quantity Per Test
A1372286A	10x Annexin V Binding Buffer	5 mL	10 mL	20 mL	2-8℃	200 μL/0.5-1.0x10⁵ cells
A1372286B	Annexin V-FITC	100 μL	250 μL	500 μL	2-8℃. Store in the dark.	5 μL/0.5-1.0x10⁵ cells
A1372286C	PI Staining Solution	100 μL	250 μL	500 μL	2-8℃. Store in the dark.	2 μL/0.5-1.0x10⁵ cells

Images

Flow Cytometry: Annexin V-FITC/PI Apoptosis Detection Kit (A1372286)

Annexin V-FITC/PI Apoptosis Detection Kit (A1372286) - Flow Cytometry
Flow cytometric analysis of U937 cells untreated (left) or treated with 4µM camptothecin (C111281) for 4 hours using Annexin V-FITC/PI Apoptosis Detection Kit (A1372286).

References

1. Jiao Sun, Yiqing Wang, Jiaxing Zhang, Jianhao Wang, Cheng Wang.  (2025)  A drug reservoir strategy for long-acting cancer therapy using morphologically transformed amphiphilic polymeric nanoparticles.  Acta Biomaterialia,      [PMID:40617494] [10.1016/j.actbio.2025.07.006]