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Annexin V Recom FITC 25Tst

 
包装: 25Tst
运保温度: Store undiluted at 4°C
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描述:

Apoptosis is a normal physiologic process which occurs during embryonic development as well as in maintenence of tissue homeostasis. The
apoptotic program is characterized by certain morphologic features, including loss of plasma membrane asymmetry and attachment,
condensation of the cytoplasm and nucleus, and internucleosomal cleavage of DNA. Loss of plasma membrane is one of the earliest features.
In apoptotic cells, the membrane phospholipid phosphatidylserine (PS) is translocated from the inner to the outer leaflet of the plasma
membrane, thereby exposing PS to the external cellular environment. Annexin V is a 35-36 kDa Ca2+ dependent phospholipid-binding
protein that has a high affinity for PS, and binds to cells with exposed PS. Annexin V may be conjugated to fluorochromes including FITC.
This format retains its high affinity for PS and thus serves as a sensitive probe for flow cytometric analysis of cells that are undergoing
apoptosis. Since externalization of PS occurs in the earlier stages of apoptosis, FITC Annexin V staining can identify apoptosis at an earlier
stage than assays based on nuclear changes such as DNA fragmentation.
FITC Annexin V staining precedes the loss of membrane integrity which accompanies the latest stages of cell death resulting from either
apoptotic or necrotic processes. Therefore, staining with FITC Annexin V is typically used in conjunction with a vital dye such as propidium
iodide (PI) or 7-Amino-Actinomycin (7-AAD) to allow the investigator to identify early apoptotic cells (PI negative, FITC Annexin V
positive). Viable cells with intact membranes exclude PI, whereas the membranes of dead and damaged cells are permeable to PI. For
example, cells that are considered viable are both FITC Annexin V and PI negative while cells that are in early apoptosis are FITC Annexin V
positive and PI negative, while cells that are in late apoptosis or already dead are both FITC Annexin V and PI positive. This assay does not
distinguish between cells that have undergone apoptotic death versus those that have died as a result of a necrotic pathway because in either
case, the dead cells will stain with both FITC Annexin V and PI. However, when apoptosis is measured over time, cells can be often tracked
from FITC Annexin V and PI negative (viable, or no measurable apoptosis), to FITC Annexin V positive and PI negative (early apoptosis,
membrane integrity is present) and finally to FITC Annexin V and PI positive (end stage apoptosis and death). The movement of cells through
these three stages suggests apoptosis. In contrast, a single observation indicating that cells are both FITC Annexin V and PI positive, in of
itself, reveals less information about the process by which the cells underwent their demise.
FITC Annexin V is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during
antibody development only or reported in the literature.


注意事项:


1.Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2.Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
3.Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
 


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