Key features and details
- Assay type: Direct
- Platform: Flow cytometer, Fluorescence microscope
- Assay time: 10 min
- Sample type: Adherent cells, Suspension cells
Product nameAnnexin V-Cy3 Apoptosis Staining / Detection Kit with SYTOX
See all Annexin V/ANXA5 kits
Sample typeAdherent cells, Suspension cells
Assay time0h 10m
Annexin V-Cy3 Apoptosis Staining / Detection Kit with SYTOX (ab14144) includes Annexin V-Cy3, SYTOX green dye, and binding buffer. It can be used in a 10 min, one-step staining procedure to detect apoptosis and necrosis. Analysis is by flow cytometry or fluorescence microscopy.
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-Cy3 and SYTOX Green dye in binding buffer, apoptotic cells show red fluorescence, dead cells show green fluorescence and live cells show little or no fluorescence.
These populations can easily be distinguished by fluorescence microscopy using FITC and rhodamine filters or by flow cytometry using the FL1 channel (Ex. 488 nm/Em. 530 nm) for SYTOX Green dye and FL2 channel for Annexin V-Cy3 (Ex. 543 nm/Em. 570 nm).
The Annexin V-Cy3 reagent contained in the kit is also available as Annexin V-Cy3 reagent ab14082.
The Annexin V apoptosis assay is based on the observation that soon after initiating apoptosis, cells 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.
PlatformFlow cytometer, Fluorescence microscope
Storage instructionsStore at +4°C. Please refer to protocols.
Components 100 tests Annexin V-Cy3 1 x 500µl Binding Buffer 1 x 50ml SYTOX Green Dye 1 x 100µl
FunctionThis protein is an anticoagulant protein that acts as an indirect inhibitor of the thromboplastin-specific complex, which is involved in the blood coagulation cascade.
Involvement in diseasePregnancy loss, recurrent, 3
Sequence similaritiesBelongs to the annexin family.
Contains 4 annexin repeats.
DomainThe [IL]-x-C-x-x-[DE] motif is a proposed target motif for cysteine S-nitrosylation mediated by the iNOS-S100A8/A9 transnitrosylase complex.
A pair of annexin repeats may form one binding site for calcium and phospholipid.
modificationsS-nitrosylation is induced by interferon-gamma and oxidatively-modified low-densitity lipoprotein (LDL(ox)) possibly implicating the iNOS-S100A8/9 transnitrosylase complex.
- Information by UniProt
- Anchorin CII
- Annexin 5
- Annexin A5
- FCCP, mitochondrial oxidative phosphorylation uncoupler (ab120081)
- Cycloheximide, Protein synthesis inhibitor (ab120093)
- SB431542, ALK inhibitor (ab120163)
- Z-VAD(OH)-FMK, Irreversible general caspase inhibitor (ab120382)
- Z-VAD(OMe)-FMK, Cell permeable, irreversible pan-caspase inhibitor (ab120487)
- Doxorubicin hydrochloride, Topoisomerase II inhibitor (ab120629)
- Dorsomorphin (Compound C), AMP-kinase inhibitor (ab120843)
- Annexin V/ANXA5-FITC Apoptosis Detection Reagent (500X) (ab14082)
- Propidium Iodide (ab14083)
- 10X Annexin V/ANXA5 Binding Buffer (ab14084)
- Annexin V-FITC Apoptosis Staining / Detection Kit (ab14085)
- Annexin V-Cy3 Apoptosis Staining / Detection Kit (ab14142)
- Calcein AM, fluorescent dye for cell viability (ab141420)
- Annexin V/ANXA5-Cy3 Apoptosis Staining / Detection Reagent (ab14143)
- Annexin V/ANXA5-Cy5 Apoptosis Staining / Detection Reagent (ab14147)
- Annexin V-Cy5 Apoptosis Staining / Detection Kit (ab14150)
- Annexin V-EGFP Apoptosis Staining / Detection Kit (ab14153)
- Annexin V/ANXA5-PE Apoptosis Staining / Detection Reagent (ab14154)
- Annexin V-PE Apoptosis Staining / Detection Kit (ab14155)
- Annexin V-PE-Cy5 Apoptosis Staining / Detection Kit (ab14159)
- Annexin V/ANXA5-Biotin Apoptosis Staining / Detection Reagent (ab14165)
ab14144 has been referenced in 5 publications.
- Du Z et al. Meningioma transcription factors link cell lineage with systemic metabolic cues. Neuro Oncol 20:1331-1343 (2018). PubMed: 29660031
- Alshatwi AA et al. The apoptotic effect of hesperetin on human cervical cancer cells is mediated through cell cycle arrest, death receptor, and mitochondrial pathways. Fundam Clin Pharmacol N/A:N/A (2012). PubMed: 22913657
- Ramesh E & Alshatwi AA Naringin induces death receptor and mitochondria-mediated apoptosis in human cervical cancer (SiHa) cells. Food Chem Toxicol 51C:97-105 (2012). PubMed: 22847135
- Pliss A et al. From the Cover: Biophotonic probing of macromolecular transformations during apoptosis. Proc Natl Acad Sci U S A 107:12771-6 (2010). PubMed: 20615987
- Honma K et al. TNFAIP3/A20 functions as a novel tumor suppressor gene in several subtypes of non-Hodgkin lymphomas. Blood 114:2467-75 (2009). Human . PubMed: 19608751