Analys av apoptos hos Docetaxel- och - DiVA

PPT - Docent Jan Lindvall Jan.Lindvall@swipnet.se

The other  2.3.6 Flow cytometric exclusion of apoptosis via Annexin V assay and TUNEL assay. 18 Double staining with Annexin-V/FITC and propidium iodide. using propidium iodide or the acriflavine-Feulgen staining procedure. Apoptosis was detected using Annexin V-FITC and propidium iodide in Panc 28 cells  MTT and Annexin V/PI assays were used to analyse cytotoxicity of KAN0439834. Immunohistochemistry staining of spleens from the two studies showed a  av MS Carro · 2016 · Citerat av 17 — subtypes. Among the predicted regulators, Annexin A2 (ANXA2) stands pCHMWS lentiviral vector (kindly provided by V. Baekelandt,. av MM DZEBO · 2014 — The cell viability can be determined by an assay using Annexin V commonly membrane, see Figure 6.6, whereas membrane staining is hard to detect for the  av H Zeng · 2018 · Citerat av 43 — Representative H&E-stained lung section shows metastasized lesion 5 μl of APC-labelled antibody against ANNEXIN V (BioLegend 640920)  21 dec.

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After taking the cells from the culture keep them on ice at all times. Prepare a stock solution of annexin V and PI in BB, so that every sample will receive 10 μL from the stock. Labeled annexin-V, with its high affinity for phosphatidylserine (PS), is a sensitive probe for Annexin V/7-amino-actinomycin staining is a convenient way to discriminate early apoptosis from late apoptosis and necrosis. Early apoptotic cells express phosphatidylserines (PS) on the outer leaflet of the plasma membrane. Annexin V, a 36-kDa calcium-binding protein, binds to PS; therefore, fluorescently labeled Annexin V can be used to detect PS that is exposed on the outside of apoptotic cells.

Annexin V and propidium iodide (PI) labeling of cells is a technique used to identify cell death, and distinguish between its different pathways: apoptosis, or programmed cell death, and necrosis. Cells undergo distinct morphological changes depending on the pathway. Annexin V/PI double staining was performed using an Annexin-V-FLUOS Staining Kit (Roche Applied Science, 11988549001) according to the manufacturer’s protocol.

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University of Florence Protocol A: Annexin V staining Protocol B: Annexin V staining with Fixable Viability Dyes Protocol C: Annexin V staining with surface and intracellular staining Introduction Annexins are a family of calcium-dependent phospholipid-binding proteins that preferentially bind phosphatidylserine (PS). The Annexin V/PI protocol is a commonly used approach for studying apoptotic cells. PI is used more often than other nuclear stains because it is economical, stable and a good indicator of cell viability, based on its capacity to exclude dye in living cells.

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no. 556420, 556419) or Annexin V-Biotin (cat. no. 556418, 556417).

In the present study, we used annexin V-propidium iodide staining to examine apoptosis and necrosis after simulated ischemia and simulated reperfusion in rat ventricular myocytes. Annexin V binds phosphatidylserine, a phosphoaminolipid thought to be externalized during apoptosis or programmed cell death. Dual staining with fluorescent Annexin V and propidium iodide (PI) has been used to discriminate apoptotic and necrotic cell death, in which Annexin V-positive/PI-negative staining is regarded as apoptosis and PI-positive staining as necrosis. Annexin V and propidium iodide (PI) labeling of cells is a technique used to identify cell death, and distinguish between its different pathways: apoptosis, or programmed cell death, and necrosis. Cells undergo distinct morphological changes depending on the pathway.
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Annexin V Staining of B1-8f/3-83κ and B1-8f/3-83κ/Mx-Cre B Cells After IFN Culture BM from control B1-8f/3-83κ and B1-8f/3-83κ/Mx-Cre mice was grown in IL-7 for 5 d. Annexin V Binding Buffer is recommended for use with Annexin V staining. Annexin V binding alone cannot differentiate between apoptotic and necrotic cells. To help distinguish between the necrotic and apoptotic cells we recommend use of our 7-amino-actinomycin D (7-AAD) solution.

2011-01-06 · Annexin V-Cy3 Apoptosis Staining / Detection Kit ab14142 is used in a 10 min, one-step staining procedure to detect apoptosis by staining phosphatidylserine molecules which have translocated to the outside of the cell membrane. Annexin V-FITC Apoptosis Staining / Detection Kit (with dead cell stain) (ab273273) is based on the observation that soon after initiating apoptosis, most cell types translocate the membrane phospholipid phosphatidylserine (PS) from the inner face of the plasma membrane to the cell surface. Add annexin V and calcium only 10 minutes before acquisition. Protocol 2: Annexin V staining experimental protocol.
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Staining cells simultaneously with FITC-Annexin V (green fluorescence) and the non-vital dye propidium iodide (red fluorescence) allows (bivariate analysis) the discrimination of intact cells (FITC-PI-), early apoptotic (FITC+PI-) and late apoptotic or necrotic cells (FITC+PI+). Annexin-V-FLUOS binds in a Ca 2+ -dependent manner to negatively charged phospholipid surfaces, and shows high specificity for phosphatidylserine. Therefore, it stains apoptotic and necrotic cells. Propidium iodide stains only the DNA of leaky necrotic cells and allows for a distinction between apoptotic and necrotic cells. Annexin V/7-AAD staining in keratinocytes. Zimmermann M(1), Meyer N. Author information: (1)Schweizerisches Institut für Allergie und Asthma Forschung (SIAF), Davos, Switzerland. maya.zimmermann@siaf.uzh.ch Annexin V/7-amino-actinomycin staining is a convenient way to discriminate early apoptosis from late apoptosis and necrosis.

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6 D–F. The granulosa cells were  Simultaneous staining of cells with AbFluor™ 488 and propidium iodide (PI) allows the discrimination of intact cells, early apoptotic and late apoptotic or necrotic  Annexin-V/PI double-staining assay. After treating with PK, cells were stained with annexin V-FITC and propidium iodide and analyzed by fluorescence  31 Oct 2018 Fluorescent images show Annexin V-FITC positive apoptotic cells and the PI positive necrotic cells. Click export.

( b ) Annexin V / 7-aminoactinomycin D-analys visar att CPB kan förbättra apoptos och The gel was stained using the Silver Stain Plus staining kit (Bio-​Rad)  Felstavar betecknar b - d Annexin V / 7AAD-färgning efter 72 timmars using the annexin V-FITC and 7-aminoactinomycin D (7AAD) staining from BD  Med tanke på att vi hittade jämförbara annexinV- och CD63-positiviteter både i MVs were set using staining controls (stainings in annexinV binding buffer).