Vybrant ® Apoptosis Assay Kit
Alexa Fluor® 488 annexin V/propidium iodide, 50 assays
Introduction
Apoptosis is a carefully regulated process of cell death that occurs as a normal part of development. Inappropriately regulated apoptosis is implicated in disease states, such as Alzheimerʼs disease and cancer. Apoptosis is distinguished from necrosis, or accidental cell death, by characteristic morphological and biochemical changes, including compaction and fragmentation of the nuclear chromatin, shrinkage of the cytoplasm, and loss of membrane asymmetry.1-5 In normal live cells, phospha tidy l serine (PS) is located on the cytoplasmic surface of the cell membrane. However, in apop totic cells, PS is translocated from the inner to the outer leaflet of the plasma membrane, thus exposing PS to the external cellular environment.6 In leukocyte apoptosis, PS on the outer surface of the cell marks the cell for recognition and phagocytosis by macro phages.7,8 The human anticoagulant, annexin V, is a 35–36 kD Ca2+-dependent phospholipid-binding protein that has a high affinity for PS.9 Annexin V labeled with a fluorophore or biotin can identify apoptotic cells by binding to PS exposed on the outer leaflet.10
The Vybrant® Apoptosis Assay Kit #2 provides a rapid and convenient assay for apoptosis. The kit contains recombinant annexin V con jugated to one of our best and brightest fluorophores, the Alexa Fluor® 488 dye, to provide the maximum sensitivity. Alexa Fluor® 488 dye is an almos t perfect spectral match to fluorescein (FITC), but it creates brighter and more photostable conjugates. In addition, the kit include s a ready-to-use solution of the red-fluorescent propidium iodid e (PI) nucleic acid binding dye. PI is impermeant to live cells and apoptotic cells, but stains dead cells with red fluorescence, binding tightly to the nucleic acids in the cell. After staining a cell population with Alexa Fluor® 488 annexin V and PI in the provided binding buffer, apoptotic cells show green fluorescence, dead cells show red and green fluorescence, and live cells show little or no fluorescence (Figure 1)
Kit Components
• Alexa Fluor® 488 annexin V (Component A), 250 µL of a solution in 25 mM HEPES,
140 mM NaCl, 1 mM EDTA, pH 7.4, plus 0.1% bovine serum albumin (BSA)
• Propidium iodide (Component B), 100 µL of a 1 mg/mL (1.5 mM) solution in dH2O
• 5X annexin-binding buffer (Component C), 15 mL of 50 mM HEPES, 700 mM NaCl,
12.5 mM CaCl2 , pH 7.4
The kit provides sufficient reagents for 50 flow cytometry assays, based on a 100 µL assay volume. Appropriate sample concentrations range from 2 × 105 to 1 × 106 cells per milliliter.
Storage and Handling
Upon receipt, store the kit at 2–6°C, protected from light. The components of the kit should be stable for at least 6 months. DO NOT FREEZE. Alexa Fluor® 488 annexin V and propidium iodide are light sensitive. These compounds may be handled in normal room light, but avoid prolonged exposure to light.
Caution: Propidium iodide is a potential mutagen; use appropriate precautions.
Experimental Protocols
We have optimized this assay using Jurkat cells treated with camptothecin to induce apoptosis. Some modifications may be required for use with other cell types.
Ordering Information
| Description | Quantity | Price(￥) | Promotion(￥) | Cat. no. | 产地 |
| Vybrant ® Apoptosis Assay Kit | 50T | 1924 | 1450(现货) | V13241 | Invitrogen |
| 5*50T | 9000 | 6500(现货) | V13241 | Invitrogen |
温馨提示：不可用于临床ZL。
资料下载：
2009060308112638.pdf
附件下载 (下载 23 次)
请 [登录] 后再下载!
VYBRANT APOPTOSIS ASSAY KIT 1 KIT
C02-HH-V13241 Vybrant Apoptosis Assay Kit #2 *Ale...
VYBRANT APOPTOSIS ASSAY KIT
Vybrant ® Apoptosis Assay Kit
VYBRANT APOPTOSIS ASSAY KIT
沪公网安备 31011502008050号
