Annexin V-Cy3™ Apoptosis Detection Kit
SIGMA/APOAC
Synonym: Red fluorescent Annexin V apoptosis assay
Product Type: Chemical
Double staining of control cells and apoptotic cells with Annexin V-Cy3 Apoptosis Detection Kit. 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 0.5-1x106 cells/ml. The cells were stained with annexin V-Cy3.18 (1 μg/ml) and carboxyfluorescein diacetate (100 μM) in binding buffer for 10 minutes at room temperature. After staining, the cells were washed with binding buffer and observed by fluorescence microscopy. Figure A: Non-induced control cells stained with CDFA. Figure B: Cells induced with staurosporine and stained with CDFA. Figure C: Non-induced control cells from Figure A after annexin V-Cy3.18 staining. Note the absence of apoptotic cells. Figure D: Cells induced with staurosporine from Figure B stained with annexin V-Cy3.18. Note that apoptotic cells are stained both with CDFA and annexin V-Cy3.18, while necrotic cells are stained with annexin V-Cy3, but not by CDFA.
| application(s) | cell analysis detection |
| detection method | fluorometric |
| packaging | pkg of 1 kit |
| Quality Level | 200  |
| shipped in | wet ice |
| storage temp. | 2-8°C |
| technique(s) | flow cytometry: suitable |
| usage | sufficient for 200 tests |
| Biochem/physiol Actions: | Sigma′s Annexin V-Cy3 kit allows detection of annexin V bound to apoptotic cells by fluorescence microscopy. The Annexin V-Cy3 kit uses the dye Cy3.18 as the fluorochrome conjugated with annexin V. By microscopy, Cy3.18 fluoresces more brightly than the FITC conjugate. The kit includes the non-fluorescent compound 6-carboxyfluorescein diacetate (6-CFDA), which enters the cell and is hydrolyzed by the esterases present in living cells to the fluorescent compound 6-carboxyfluorescein, indicating that the cells are viable. This combination allows the differentiation among early apoptotic cells (annexin V positive, 6-CFDA positive), necrotic cells (annexin V positive, 6-CFDA negative), and viable cells (annexin V negative, 6-CFDA positive). |
| 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. • 6-Carboxyfluorescein diacetate secondary dye is included with the kit to differentiate apoptotic cells from viable and necrotic cells. |
| Legal Information: | Cy3 is a trademark of Cytiva |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352207 |
| Components | 6-Carboxyfluorescein diacetate 10 mg; Annexin V Cy3.18 Conjugate from human placenta, buffered aqueous solution 10 μg |