Cells were grown on coverslips coated with PDL/laminin and were stained for protein aggresomes at 9 days post lentiviral infection for CTRL or KDSR KO or 18hr post treatment with vehicle or 3μM MG132. Protein aggresomes were detected using the PROTOEOSTAT Aggresome detection kit (Enzo Life Sciences #ENZ-51035), which contains a dye that fluoresces upon intercalation into the cross-beta spine of protein structures found in misfolded and aggregated proteins. For aggresome staining, cells were fixed in 4% PFA, washed in PBS, and permeabilized in 0.5% TXBon ice with gentle shaking for 30 min. Cells were washed in PBS following permeabilization and were subsequently incubated with the Aggresome Detection Reagent diluted 1:2000 in PBS for 45 min at room temperature protected from light. Cells were washed with PBS after staining and coverslips were mounted to microscope slides using ProLong Gold Antifade Mountant containing DAPI (Thermofisher #P36941). Cells were imaged using the Zeiss LSM700 confocal microscope and Zen Black 2012 Software.
Protoeostat aggresome detection kit
Manufactured by Enzo Life Sciences
The PROTOEOSTAT Aggresome detection kit is a laboratory tool used for the detection and visualization of aggresomes within cells. Aggresomes are cellular structures that form when misfolded proteins accumulate in the cytoplasm. The kit provides the necessary reagents and protocols to facilitate the identification and analysis of aggresomes in biological samples.
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2 protocols using protoeostat aggresome detection kit
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Protein Aggresomes Detection in Cell Lines
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Protein Aggresomes Detection in Cell Lines
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Cells were grown on coverslips coated with PDL/laminin and were stained for protein aggresomes at 9 days post lentiviral infection for CTRL or KDSR KO or 18hr post treatment with vehicle or 3μM MG132. Protein aggresomes were detected using the PROTOEOSTAT Aggresome detection kit (Enzo Life Sciences #ENZ-51035), which contains a dye that fluoresces upon intercalation into the cross-beta spine of protein structures found in misfolded and aggregated proteins. For aggresome staining, cells were fixed in 4% PFA, washed in PBS, and permeabilized in 0.5% TXBon ice with gentle shaking for 30 min. Cells were washed in PBS following permeabilization and were subsequently incubated with the Aggresome Detection Reagent diluted 1:2000 in PBS for 45 min at room temperature protected from light. Cells were washed with PBS after staining and coverslips were mounted to microscope slides using ProLong Gold Antifade Mountant containing DAPI (Thermofisher #P36941). Cells were imaged using the Zeiss LSM700 confocal microscope and Zen Black 2012 Software.
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