Annexin V-FITC Apoptosis Detection Kit
SIGMA/APOAF
Synonym: Annexin V-FITC; Apoptosis probe FITC
Product Type: Chemical
Detection of apoptosis by fluorescence microscopy in Jurkat cells treated with staurosporine. Apoptosis was induced in Jurkat cells by 1 μg/ml staurosporine for 2 hours. The cells were washed with PBS and suspended at a concentration of 0.5-x106 cells/ml. The cell suspension was applied to a poly-L-lysine-coated slide and allowed to adhere for 10 minutes. The cells were then washed with binding buffer (10mM HEPES, pH 7.5, 140 mM NaCl, 2.5 mM CaCl2). The cells were stained with annexin V-FITC (2 μg/ml) for 10 minutes at room temperature. After staining, the cells were washed with binding buffer and observed by fluorescence microscopy.
Flow Cytometry 1×106 Jurkat cells were incubated with 1 μg/mL staurosporine followed by 5 μL of Annexin V-FITC Conjugate and 10 μL of Propidium Iodide Solution. Purple histogram: sample with staurosporine ; Green histogram: sample without staurosporine. (Staurosporine induce apoptosis).
| application(s) | cell analysis detection |
| detection method | fluorometric |
| packaging | pkg of 1 kit |
| Quality Level | 300  |
| shipped in | wet ice |
| storage temp. | 2-8°C |
| technique(s) | flow cytometry: suitable |
| usage | (20 tests) |
| Application: | Annexin V-FITC Apoptosis Detection Kit was used: • in staining of LNCaP prostate cancer cells for measuring the G. lucidum extracts activity during the treatment of prostate cancer. • for tumor cell labelling to study the inhibitory activity of DBP-maf (Vitamin D binding protein-macrophage activating factor) on prostate tumor cells. • for indirect measurement of flippase activity. |
| Features and Benefits: | Detects apoptosis earlier in the process than DNA-based assays such as TUNEL. • Rapid labeling of cells. Cell staining takes only 10 minutes. • No cell fixation or processing required, reducing the detection time and allowing the cells to be used for further study. • Propidium iodide secondary dye is included with the kit to differentiate apoptotic cells from viable and necrotic cells. |
| General description: | Annexin V-FITC kit allows fluorescent detection of annexin V bound to apoptotic cells and quantitative determination by flow cytometry. The AnnexinV-FITC kit uses annexin V conjugated with fluorescein isothiocyante (FITC) to label phosphatidylserine sites on the membrane surface. The kit includes propidium iodide (PI) to label the cellular DNA in necrotic cells where the cell membrane has been totally compromised. This combination allows the differentiation among early apoptotic cells (annexin V positive, PI negative), necrotic cells (annexin V positive, PI positive), and viable cells (annexin V negative, PI negative). |
| Other Notes: | Allow all components to reach room temperature before use. |
| RIDADR | NONH for all modes of transport |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352207 |
| Components | APOAFA |