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Annexin V-PE
Product overview:
Annexin V-PE
Catalog Number: A6065S, A6065L
Product Size: 0.1 mL, 1 mL
Storage
Store at 4℃ and protect
from light. Do not freeze. Expiration date marked on the outer packing.
Spectral Characteristics
Annexin
V-PE: Ex/Em=488/578 nm
Description
Annexin V (membrane-linked protein-V)
is a Ca2+-dependent phospholipid-binding protein with a molecular
weight of 35-36 KD that binds selectively to phosphatidylserine (PS). PS is
mainly distributed on the inner side of the cell membrane, i.e., the side
adjacent to the cytoplasm. In the early stages of apoptosis, different cell
type exocytose phosphatidylserine to the cell surface and are exposed to the
extracellular environment. At this point, the use of fluorescent protein
PE-tagged Annexin V, Annexin V-PE, in combination with the outside-out PS
allows flow cytometry to directly detect the outside-out PS, an important
feature of apoptosis. Normal cells are not stained by Annexin V-PE, cells
undergoing apoptosis or necrosis are stained by Annexin V-PE. Annexin V-PE can
be used in combination with partially non-permeable nuclear dyes (7-AAD/PI) to differentiate cells in different periods of apoptosis.
Notes
2. To reduce the process of apoptosis, incubation can be done on ice, but the incubation time should be extended to at least 30 min.
3. Because apoptosis is a rapid process, it is recommended that samples be analyzed within 1 h of staining.
4. For adherent cells, digestion is a critical step. If there are floating cells during the induction of apoptosis in the adherent cells, collect the floating cells and the walled cells and combine them for staining. Be careful when handling walled cells and try to avoid artificial damage to the cells. If the digestion time of trypsin is too short, the cells need to be blown hard to fall off, which will easily cause damage to the cell membrane and excessive intake of PI; if the digestion time is too long, the cell membrane will also be easily damaged, and even affect the binding of phosphatidylserine and Annexin V-PE on the cell membrane. When digestion, trypsin is spread all over the bottom of the well plate, and then the trypsin is fully contacted with the cells when shaken gently, and then most of the trypsin is poured off, and the remaining small amount of trypsin is used to digest for a period of time, which can be terminated when the space between cells increases and the bottom of the bottle is flowery. Try not to use EDTA in the digestion solution, EDTA will affect the binding of Annexin V to PS.
5. After digestion with trypsin, it is recommended that the walled cells be stained after about 30 min of recovery in optimal culture conditions and medium to avoid false positives.
6. To avoid cell loss when washing cells, use a large Tip tip over a small Tip tip when aspirating.
7. The optimal concentration of the dye to be used is determined by the specific experimental requirements.
8. There are quenching problems with fluorescent dyes. Please avoid light to slow down the fluorescence quenching.
9. For your safety and health, please wear lab coats and disposable gloves.
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