Annexin V-FITC Apoptosis Kit Plus

Antigen: Annexin A5 (ANXA5)

Synonyms: PP4, ANX5, ENX2, Anx5, R74653, LC5, MGC93655, ANXA5, MGC126971, anx, anx5, ANX V, anxa5, cb989, wu:fa98f06, wu:fj10f10, MGC89158

Conjugate: FITC

Application: Flow Cytometry (FACS), Fluorescence Microscopy (FM)

Catalog no.: ABIN411998

Additional Information

Alternative name: Annexin V

Description: The Annexin V-FITC Apoptosis Detection Kit Plus is based on the observation that soon after initiating apoptosis, most cell types translocate the membrane phospholipid phosphatidylserine (PS) from the inner face of the plasma membrane to the cell surface. Once on the cell surface, PS can easily be detected by staining with a fluorescent conjugate of Annexin V, a protein that has a strong natural affinity for PS. The one-step staining procedure takes only 10 minutes. In addition, the assay can be directly performed on live cells, without the need of fixation. The Annexin V-FITC Apoptosis Detection Kit Plus includes annexin V-FITC, SYTOX green dye, and binding buffer. The SYTOX green dye is impermeant to live cells and apoptotic cells, but stains necrotic cells with intense green fluorescence by binding to cellular nucleic acids. After staining a cell population with annexin V-FITC and SYTOX Green dye in the provided binding buffer, apoptotic cells show green fluorescence, dead cells show a higher level of green fluorescence and live cells show little or no fluorescence. These populations can easily be distinguished using a flow cytometry with the 488 nm line of an argon-ion laser for excitation. Both annexin V-FITC and SYTOX Green dye emit green fluorescence that can be detected in the FL1 channel, freeing the other channels for the addition of other probes in multi-color labeling experiments.

Application Details

Protocol: 1. Induce apoptosis by desired method. Concurrently incubate a control culture without induction. 2. Collect 1-5 x 105 cells by centrifugation. 3. Resuspend cells in 500 µl of 1X Binding Buffer. 4. Add 5 µl of Annexin V-FITC and 1 µl of SYTOX Green dye

Application Notes: Note: Thaw the SYTOX Green dye in room temperature before use. 5. Incubate at room temperature for 5-10 min in the dark. 6. Analyze the stained cells by flow cytometry (Ex = 488 nm, Em = 530 nm). The cell population should separate into three groups: live cells with only a low level of fluorescence, apoptotic cells with moderate green fluorescence and necrotic cells with high-intensity green fluorescence. For adherent cells, gently trypsinize and wash cells once with serum-containing media before incubation with Annexin V-FITC and SYTOX dye.

Restrictions: For Research Use only