Testing Data
(Autophagy induction in Jurkats measured by flow cytometry using Autophagy Probe, RedJurkat cells were either untreated for basal conditions (red), treated with 0.5 muM Rapamycin for 18 hours (blue), or treated with 0.5 muM Rapamycin for 18 hours and 10 nM Bafilomycin A for 2 hours (orange) to induce autophagy. After staining with Autophagy Probe, Red for 30 minutes, cells were washed and analyzed on the ZE5 Cell Analyzer using the 561 nm laser and 640/20 filter)
Autophagy Assay Assay Kit
Autophagy Assay, Red Detection Kit
Applications
Flow Cytometry, Functional Assay
Synonyms
Autophagy Assay; Red Detection Kit; Autophagy Assay Kit; Red; Autophagy Assay assay kit
Applicable Applications for Autophagy Assay assay kit
Flow Cytometry (FC/FACS)
Preparation and Storage
Store the unopened kit (and each unopened component) according to the storage instructions on each component label.
Store the Autophagy Probe, Red at -20 degree C. Once reconstituted in DMSO, the Autophagy Probe, Red stock should be stored at -20 degree C for 6 months, protected from light.
Avoid repeated freezing and thawing.
Store the Autophagy Probe, Red at -20 degree C. Once reconstituted in DMSO, the Autophagy Probe, Red stock should be stored at -20 degree C for 6 months, protected from light.
Avoid repeated freezing and thawing.
Testing Data
(Autophagy induction in Jurkats measured by flow cytometry using Autophagy Probe, RedJurkat cells were either untreated for basal conditions (red), treated with 0.5 muM Rapamycin for 18 hours (blue), or treated with 0.5 muM Rapamycin for 18 hours and 10 nM Bafilomycin A for 2 hours (orange) to induce autophagy. After staining with Autophagy Probe, Red for 30 minutes, cells were washed and analyzed on the ZE5 Cell Analyzer using the 561 nm laser and 640/20 filter)
Testing Data
(Autophagy induction in Jurkats measured by flow cytometry using Autophagy Probe, RedJurkat cells were either untreated for basal conditions (red), treated with 0.5 muM Rapamycin for 18 hours (blue), or treated with 0.5 muM Rapamycin for 18 hours and 10 nM Bafilomycin A for 2 hours (orange) to induce autophagy. After staining with Autophagy Probe, Red for 30 minutes, cells were washed and analyzed on the ZE5 Cell Analyzer using the 561 nm laser and 640/20 filter)
Testing Data
(Autophagy induction in Jurkats measured by flow cytometry using Autophagy Probe, RedJurkat cells were either untreated for basal conditions (red) or incubated in HBSS for 2 hours and 10 nM Bafilomycin A for 2 hours (blue) to induce autophagy. After staining with Autophagy Probe Red for 30 minutes, cells were washed and analyzed on the ZE5 Cell Analyzer using the 561 nm laser and 640/20 filter.)
Testing Data
(Autophagy induction in Jurkats measured by flow cytometry using Autophagy Probe, RedJurkat cells were either untreated for basal conditions (red) or incubated in HBSS for 2 hours and 10 nM Bafilomycin A for 2 hours (blue) to induce autophagy. After staining with Autophagy Probe Red for 30 minutes, cells were washed and analyzed on the ZE5 Cell Analyzer using the 561 nm laser and 640/20 filter.)
Related Product Information for Autophagy Assay assay kit
Summary: Autophagy Assay, Red Detection Kit allows for the detection and monitoring of in vitro development of autophagy in living cells.
Autophagy is a conserved lysosomal recycling process by which cells break down their own components such as proteins, lipids and carbohydrates. Autophagy plays a critical role in maintaining homeostasis by preventing the accumulation of damaged organelles by disassembling unnecessary or dysfunctional cells and cellular components (Mizushima et al 2011). Autophagy occurs at low levels in the cell under normal conditions and can be rapidly upregulated during times of starvation or stress. Such degradation activities serve to provide nutrients (amino acids, nucleotides, fatty acids, etc.) and energy during periods of elevated bioenergetic demands (Mizushima et al 2011, Levine et al 2008). Another function of autophagy is to assist with the detection and destruction of intracellular pathogens (viruses, bacteria and parasites) (Levine et al 2011). Dysregulation of autophagy has been associated with many disease states including cancer, infection and degenerative diseases (Levine et al 2008). Autophagy is a dynamic process typically divided into three stages. During stage one, cytoplasmic components targeted for degradation are sequestered within a double-membrane phagopore (also called the isolation membrane). This results in the formation of a double-membrane vesicle called the autophagosome. During stage two, the autophagosome fuses with the lysosome to form the autolysosome. Degradation of the autophagosomal contents occurs during stage three (Mizushima et al 2011, Hundeshagen et al 2011).
Test Principle: Autophagy Probe, Red is a cell-permeant aliphatic molecule that fluoresces brightly when inserted in the lipid membranes of autophagosomes and autolysosomes. Autophagy Probe, Red can be readily detected by flow cytometry with optimal excitation at 590 nm and peak emission at 620 nm (ZE5 Cell Analyzer settings, 561 nm laser and 615/24 or 640/20 filter).
Autophagy is a conserved lysosomal recycling process by which cells break down their own components such as proteins, lipids and carbohydrates. Autophagy plays a critical role in maintaining homeostasis by preventing the accumulation of damaged organelles by disassembling unnecessary or dysfunctional cells and cellular components (Mizushima et al 2011). Autophagy occurs at low levels in the cell under normal conditions and can be rapidly upregulated during times of starvation or stress. Such degradation activities serve to provide nutrients (amino acids, nucleotides, fatty acids, etc.) and energy during periods of elevated bioenergetic demands (Mizushima et al 2011, Levine et al 2008). Another function of autophagy is to assist with the detection and destruction of intracellular pathogens (viruses, bacteria and parasites) (Levine et al 2011). Dysregulation of autophagy has been associated with many disease states including cancer, infection and degenerative diseases (Levine et al 2008). Autophagy is a dynamic process typically divided into three stages. During stage one, cytoplasmic components targeted for degradation are sequestered within a double-membrane phagopore (also called the isolation membrane). This results in the formation of a double-membrane vesicle called the autophagosome. During stage two, the autophagosome fuses with the lysosome to form the autolysosome. Degradation of the autophagosomal contents occurs during stage three (Mizushima et al 2011, Hundeshagen et al 2011).
Test Principle: Autophagy Probe, Red is a cell-permeant aliphatic molecule that fluoresces brightly when inserted in the lipid membranes of autophagosomes and autolysosomes. Autophagy Probe, Red can be readily detected by flow cytometry with optimal excitation at 590 nm and peak emission at 620 nm (ZE5 Cell Analyzer settings, 561 nm laser and 615/24 or 640/20 filter).
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Product Notes
The Autophagy Assay (Catalog #AAA225618) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's Autophagy Assay can be used in a range of immunoassay formats including, but not limited to, Flow Cytometry (FC/FACS). Researchers should empirically determine the suitability of the Autophagy Assay for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Autophagy Assay, Assay Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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