Propidium Iodide Staining Apoptosis – High affinity and specificity annexin v fitc/pi apoptosis kit e ck a211 at elabscience.com, My 3 step approach to gating annexin v data appropriately, Isobaric tags for relative and absolute quantitation based quantitative proteomic analysis of x linked inhibitor of apoptosis and h2ax in etoposide induced renal cell carcinoma apoptosis, Enhancement in efferocytosis of oxidized low‑density lipoprotein induced apoptotic raw264.7 cells through sirt1 mediated autophagy, Cisplatin induced renal cell apoptosis: caspase 3 dependent and independent pathways, Resistance to apoptosis in ctll 2 cells constitutively expressing c myb is associated with induction of bcl 2 expression and myb dependent regulation of bcl 2 promoter activity
You are now leaving the BD Bioscience website. The website you are about to visit is operated by a third party. A link to this website does not constitute any representation or warranty as to the products or services offered on the third party website and is provided only to provide convenient access to the third party website and for no other purpose. Do you want to continue?
Flow cytometric analysis of FITC annexin V staining. Jurkat cells (human T-cell leukemia; ATCC TIB-152) were left untreated (upper panel) or treated for 4 h with 12 µM camptothecin (lower panel). Cells were incubated with FITC annexin V in buffer containing propidium iodide (PI) and analyzed by flow cytometry. Untreated cells were predominantly FITC-annexin V and PI negative, indicating that they were viable and did not undergo apoptosis. After 4 h of treatment (bottom panel), there were mainly two populations of cells: cells that were viable and not undergoing apoptosis (FITC annexin V and PI negative) and cells that underwent apoptosis (FITC annexin V positive and PI negative). A small population of cells appeared to be FITC-annexin V and PI positive, indicating that they were in the final stages of apoptosis or had already died.
Propidium Iodide Staining Apoptosis
Use for research only. Not intended for use in diagnostic or therapeutic procedures. Although not mandatory, these products are manufactured in accordance with good manufacturing practices
Apoptosis Analysis By Propidium Iodide Staining And Flow Cytometry. A,…
Any unauthorized use of products without the express written permission of Becton, Dickinson & Company is strictly prohibited.
Apoptosis is a normal physiological process that occurs during embryonic development as well as in the maintenance of tissue homeostasis. The apoptotic program is characterized by certain morphological features, including plasma membrane heterogeneity and loss of attachment, cytoplasmic and nuclear condensation, and internucleosomal cleavage of DNA. One of the earliest symptoms is damage to the plasma membrane. In apoptotic cells, the membrane phospholipid phosphatidylserine (PS) is translocated from the interior of the plasma membrane to the outer leaflet, exposing PS to the extracellular environment. Annexin V is a 35–36 kDa Ca2+-dependent phospholipid-binding protein with high affinity for PS and binds to cells with exposed PS. Annexin V can be conjugated to a fluorochrome with FITC. This form retains its high affinity for PS and thus serves as a sensitive probe for flow cytometric analysis of cells undergoing apoptosis. Because PS externalization occurs at an early stage of apoptosis, FITC annexin V staining can detect apoptosis at an earlier stage than assays based on molecular changes such as DNA fragmentation.
FITC annexin V staining precedes the loss of membrane integrity that accompanies later stages of cell death due to apoptotic or necrotic processes. Therefore, FITC annexin V staining is usually used in conjunction with a key dye such as propidium iodide (PI) or 7-amino-actinomycin (7-AAD) to allow investigators to identify early apoptotic cells (PI negative, FITC annexin V positive. ). Viable cells with intact membranes exclude PI, while dead and damaged cell membranes are permeable to PI. For example, cells considered viable are FITC annexin V and PI negative; Cells showing early apoptosis are FITC annexin V positive and PI negative; and late apoptosis or already dead cells are FITC annexin V and PI positive. This assay does not distinguish cells that have died by a necrotic pathway from those that have died apoptotically, because in both cases dead cells stain with both FITC annexin V and PI. However, when apoptosis is measured over time, cells can be tracked from FITC annexin V and PI negative (viable or measurable apoptosis) to FITC annexin V positive and PI negative (early apoptosis, membrane integrity) and finally FITC annexin V. and PI positive (end-stage apoptosis and death). Movement of cells through these three stages is indicative of apoptosis. In contrast, the single observation that cells are both FITC-annexin V and PI positive reveals little information about the process by which the cells were destroyed.
FITC Annexin V is used to measure the percentage of cells in a population that are actively undergoing apoptosis. It is based on the property of cells to lose membrane heterogeneity in the early stages of apoptosis. In apoptotic cells, the membrane phospholipid phosphatidylserine (PS) is translocated from the inner leaflet of the plasma membrane to the outer leaflet, exposing PS to the external environment. Annexin V is a calcium-dependent phospholipid-binding protein with high affinity for PS and is useful for identifying apoptotic cells with exposed PS. Propidium iodide (PI) is a standard flow cytometric viability assay and is used to distinguish viable from non-viable cells. Viable cells with intact membranes exclude PI, while dead and damaged cell membranes are permeable to PI. Cells that are positive for FITC annexin V and negative for PI undergo apoptosis. Cells that stain positive for both FITC annexin V and PI are either in the final stages of apoptosis, undergoing necrosis, or have already died. Cells stained negative for both FITC annexin V and PI are alive and do not undergo measurable apoptosis.
The Images Of Apoptotic Cells By Acridine Orange/propidium Iodide…
3. 10X Annexin V Binding Buffer (Part No. 51-66121E): 0.1 M Hepes/NaOH (pH 7.4), 1.4 M NaCl, 25 mM CaCl2. For a 1X working solution, dilute 1 part 10X Annexin V Binding Buffer with 9 parts distilled water.
To obtain positive control staining with FITC annexin V and/or FITC annexin V and PI, a cell line that can be easily induced into apoptosis should be used. It is important to note that baseline levels of apoptosis and necrosis vary widely among populations. Thus, even if apoptosis is not induced, most cell populations will contain a small percentage of cells positive for apoptosis (FITC annexin V positive, PI negative or FITC annexin V positive, PI positive).
Crude populations are used to determine baseline levels of apoptotic and dead cells. The percentage of cells undergoing apoptosis is then determined by subtracting the percentage of apoptotic cells in the untreated population from the percentage of apoptotic cells in the treated population. Because cell death is a possible outcome for cells undergoing apoptosis, cells in the late stages of apoptosis are membrane damaged and stain positive for PI as well as FITC annexin V. Thus, the assay does not differentiate cells that are already apoptotic. Cell death and those that have died by a necrotic pathway, as in both cases dead cells stain with both FITC annexin V and PI.
GLOBAL – Consult the manufacturer’s instructions for use and relevant user manuals and technical data sheets before using these products as described.
Annexin V Propidium Iodide Staining For Detecting…
Comparisons are made, where applicable, with older BD technologies, manual methods or general performance claims. Comparisons are not made with non-BD technologies unless otherwise noted. You are now leaving the BD Bioscience website. The website you are about to visit is operated by a third party. A link to this website does not constitute any representation or warranty as to the products or services offered on the third party website and is provided only to provide convenient access to the third party website and for no other purpose. Do you want to continue?
Flow cytometric analysis of cells after induction of apoptosis. Jurkat leukemia cells were left untreated (upper left and lower left panels) or treated for 5 h (middle upper and middle lower panels) or 12 h (upper right and lower right panels) with an anti-human Fas antibody (clone DX2, Cat. was done 555670) and protein G (addition of protein G enhances the ability of DX2 to induce apoptosis, possibly by binding Fas). Cells were incubated with SAv-FITC (cat no. 554060) in annexin V-biotin (cat no. 556417) followed by propidium iodide (PI) staining solution (cat no. 556463). Cells were then analyzed by flow cytometry. Untreated cells were predominantly Annexin V-biotin and PI negative, indicating that they were viable and did not undergo apoptosis. After 5 h of DX2 treatment, there were two populations of cells: cells that underwent apoptosis (annexin V-biotin positive and PI negative) and cells that were viable and did not undergo apoptosis (annexin V-biotin and PI negative). After 12 h of DX2 treatment, three cell populations were identified: cells that had already died or were in the final stages of apoptosis (annexin V-biotin and PI positive), cells undergoing apoptosis (annexin V-biotin positive and PI negative), and cells that were viable. were and did not undergo apoptosis (annexin V-biotin and PI negative).
Prodium iodide (PI) staining solution can be used to assess plasma membrane (PM) integrity in annexin V apoptosis assays. PI is a fluorescent dye that stains DNA. It does not cross the PM of viable or early apoptotic cells, as they maintain PM integrity. In contrast, cells in the late stages of apoptosis or already dead have lost PM integrity
Resistance To Apoptosis In CTLL 2 Cells Constitutively Expressing C Myb Is Associated With Induction Of BCL 2 Expression And Myb Dependent Regulation Of Bcl 2 Promoter Activity, Calcein+/PI As An Early Apoptotic Feature In Leishmania, Comparison Of Cell Based Assays To Quantify Treatment Effects Of Anticancer Drugs Identifies A New Application For Bodipy L Cystine To Measure Apoptosis, An Equivalence Of Prokaryotic Pore Forming Proteins Of Plasmodium Triggers Cellular Dysfunction Responsible For Malaria Pathogenesis, Propidium Iodide, Detection Of Apoptosis Using Flow Cytometry After Annexin…, DNA Fragmentation And Apoptosis, A New Four‐color Flow Cytometric Assay To Detect Apoptosis In Lymphocyte Subsets Of Cultured Peripheral Blood Cells, Cell Viability & Apoptosis