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CYTO-ID® Autophagy detection kit - ENZ - Enzo Life Sciences
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CYTO-ID ® Autophagy detection kit A no-transfection, quantitative assay for monitoring autophagy in live cells. ENZ 50 tests No transfection required Proprietary dye includes titratable moieties specific for selectively staining autophagic vesicles Protocol validated with known inhibitors and activators of autophagic activity Rapidly quantifies autophagy in native heterogeneous cell populations Eliminates need for time and effort-consuming transfection efficiency validation required with LC3-GFP transfection Selective and comprehensive staining, allows measurement master thesis supplier involvement differentiation between autophagic flux and autophagolysosome accumulation Negligible staining of lysosomes reduces background seen with other dyes Facilitates high-throughput screening of activators and inhibitors of autophagy.
CYTO-ID ® Autophagy Detection Kit measures autophagic vacuoles and monitors autophagic flux in lysosomally inhibited live cells using a novel dye that selectively labels accumulated autophagic vacuoles. The dye has been optimized through the identification of titratable functional moieties that allow for minimal staining of lysosomes while exhibiting bright fluorescence upon incorporation into pre-autophagosomes, autophagosomes, and autolysosomes autophagolysosomes.
The assay offers a rapid and master thesis supplier involvement approach to monitoring autophagy in live cells without the need for cell transfection. Mechanism of Action The probe is a cationic amphiphilic tracer CAT dye that rapidly partitions into cells in a similar manner as drugs that induce phospholipidosis.
Careful selection of titratable functional moieties on the dye prevents its accumulation within lysosomes, but enables labeling of vacuoles associated with the autophagy pathway. CYTO-ID ® Autophagy Detection Kit. Time-saving, rapid and comprehensive labeling of autophagic vacuoles without transfection. For the purpose of demonstrating advantages of CYTO-ID ® Green detection reagent, HeLa cells were first transfected with RFP-LC3 expression vector, treated with 10 µM Tamoxifen overnight, then stained with CYTO-ID ® Green detection reagent.
Panel A: Green signal indicating CYTO-ID ® Green staining of autophagic vesicles; Panel B: RFP-LC3 expression red in a subset of successfully transfected cells; Panel C: Composite image, showing CYTO-ID ® Green dye-labeled vesicles co-localize with LC3, a specific marker of autophagosomes. Profile autophagy without transfection. Figure 1A: CHO cells stably expressing GFP-LC3 transfected cell lines results in relatively poor baseline separation of control-vs-starved cell populations, making quantification of autophagy difficult.
Figure adapted from Shvets E, Fass E, Elazar Z. Figure 1B: The CYTO-ID ® Autophagy Detection Kit specifically labels autophagic vacuoles independent of LC3 protein and eliminates the need for transfection. HeLa cells were subjected to starvation and recovery and then labeled with CYTO-ID ® Green detection reagent. The dye enables clear detection and quantification of autophagic and pre-autophagic vacuoles that directly correlates to induction of autophagy. Visualization of autophagic accumulation and autophagic flux.
Autophagic vacuole accumulation and flux are both detected by CYTO-ID ® Autophagy Green dye as observed by fluorescence microscopy. HeLa cells were mock-induced with 0. After treatment, cells were incubated with CYTO-ID ® Green Detection reagent for 10 min at 37°C and then washed with assay buffer. Nuclei were counter-stained in blue with Hoechst dye. Master thesis supplier involvement background resulting from non-specific lysosomal staining.
CYTO-ID ® Green dye eliminates background staining of lysosomes seen with other lysosomotrophic dye-based assays that utilize monodansylcadaverine MDC bottom panel.
The CYTO-ID ® Autophagy kit eliminates the master thesis supplier involvement for a nm UV laser for live cell analysis, and is compatible for use with Hoechst dyes for co-labeling in microscopy applications. Overnight incubation of HepG2 cells with Rapamycin, an inhibitor of mTOR kinase, results in an increase in CYTO-ID ® dye signal.
Flow cytometry-based profiling of autophagy with CYTO-ID® Autophagy Detection Kit: Control red-lined peak uninduced and 10uM Tamoxifen ALX treated blue-filled peak Jurkat cells T-cell leukemia were used. After 18 hours treatment, cells were loaded with CYTO-ID® Green Detection Reagent, then analyzed without washing by flow cytometry. Results are presented by histogram overlays. Control cells were stained as well but mostly display low fluorescence.
In the samples treated with 10uM Tamoxifen for 18 hours, CYTO-ID® Green dye signal master thesis supplier involvement about 2-fold, indicating that Tamoxifen causes an increase in autophagy in Jurkat cells. Schematic depiction of autophagy.
Cytosolic material is sequestered by an expanding membrane sac, the phagophore, resulting in the formation of a double-membrane vesicle, an autophagosome. The outer membrane of the autophagosome subsequently fuses with the lysosome, master thesis supplier involvement, and the internal material is degraded in the autolysosome.
Various regulators of autophagy are also depicted in the diagram, master thesis supplier involvement. ABCE1 Regulates RNase L-Induced Autophagy during Viral Infections : B. Ramnan, et al, master thesis supplier involvement. Advanced Maternal Age Deteriorates the Developmental Competence of Vitrified Oocytes in Mice : J. Lee, et al. Aggregated Tau-PHF6 VQIVYK Potentiates NLRP3 Inflammasome Expression and Autophagy in Human Microglial Cells : C. Panda, et al. Alpha-1 antitrypsin master thesis supplier involvement heme-induced endothelial cell inflammatory activation, autophagy dysfunction and death : K.
Madyaningrana, master thesis supplier involvement, et al. Alpha1-antitrypsin counteracts heme-induced endothelial cell inflammatory activation, autophagy dysfunction and death : K. Areca nut extract ANE inhibits the progression of hepatocellular carcinoma cells via activation of ROS production and activation of autophagy : P.
Wei, et al. Acharya, et al. Autophagy-Mediated Activation of Mucosal-Associated Invariant T Cells Driven by Mesenchymal Stem Cell-Derived IL : G. Ye, et al. Cynaroside protects the blue light-induced retinal degeneration through alleviating apoptosis and inducing master thesis supplier involvement in vitro and in vivo : J.
H Feng, master thesis supplier involvement al. Gαq activation modulates autophagy by promoting mTORC1 signaling : S. Cabezudo, et al.
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CYTO-ID ® Autophagy Detection Kit measures autophagic vacuoles and monitors autophagic flux in lysosomally inhibited live cells using a novel dye that selectively labels accumulated autophagic vacuoles. The dye has been optimized through the identification of titratable functional moieties that allow for minimal staining of lysosomes while exhibiting bright fluorescence upon incorporation Detroit Office FEV Consulting Inc. Glenmeade Lane, Auburn Hills, MI Phone +1 Dubai Office FEV Asia GmbH (Branch) blogger.com No. Master thesis supplier involvement. Famous nyu essay essay questions for general psychology, professional business plan proofreading for hire uk stratfield saye essayist better write essay. Popular dissertation results editor services ca, essay contract law atiyah
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