Speck formation assay
WebJan 1, 2024 · Here, we present two more methods to detect ASC speck formation: (1) Assessment of ASC speck formation by flow cytometry; and (2) Chemical cross-linking of ASC followed by immunoblotting. These ... WebThe presence of ASC::GFP positive cells and localization of fluorescent ASC specks can be determined by using time-lapse confocal or high-resolution fluorescence microscopy. …
Speck formation assay
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WebJun 11, 2024 · A high-throughput assay based on fluorescence anisotropy has been used to screen for inhibitors of the formation of ASC PYD filaments and its pH dependence . Here we developed an in vitro assay for the detection of the polymerization of ASC PYD using microscale thermophoresis (MST). MST detects movement along a temperature gradient … WebOct 1, 2024 · The protein concentration was quantified by the Bradford method using the Bio-Rad protein assay kit (Bio-Rad, Hercules, CA, USA). The protein samples were subjected to SDS-PAGE (10~14% gel) and transferred to the PVDF membrane (Merck Millipore). ... This ASC speck formation in LPS/S1P-treated BMDMs was attenuated by suppressing S1P 2 …
WebDec 13, 2024 · After 24 h, HEK293 cells were harvested to analyze the speck formation by flow cytometry and confocal microscopy as well as the oligomerization formation by … WebInflammasome assembly results in the formation of a large intracellular protein scaffold driven by the oligomerization of the adaptor protein apoptosis-associated speck-like …
WebHence, ASC speck formation can be used as a simple upstream readout for inflammasome activation. Here, we describe a method for analyzing inflammasome activation by ASC … WebMar 17, 2024 · Structure-guided biochemical and functional assays show that both NLRP1 binding and enzymatic activity are required for DPP9 to suppress NLRP1 in human cells. Together, our data reveal the...
WebApr 11, 2024 · The immunofluorescence image showed that a little of ASC speck are observed in MEECs stimulated by LPS, while a large number of ASC spots are observed in MEECs stimulated by LPS plus nigericin (Fig. 9-A,B). We found that TFC-contained serum treatment reduced the formation of ASC speck. Download : Download high-res image …
WebLive Content Imaging Multiplexed, Quantitative, Kinetic Assays enabled by IncuCyte™ Reagents and IncuCyte ZOOM®, Essen BioScience, SLAS, 12-Jan-13, Orlando, FL USA Download Simplifying high throughput 3D spheroid growth and shrinkage assays using live content imaging Download fortnite season chapter 4WebFeb 25, 2024 · Apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC) is an adaptor protein that is essential for the activation of several … dinkydoodle airbrush problemsWebDec 2, 2014 · Our results indicate that the ASC speck formation is based on specific homophilic but not heterophilic interactions by PYD and CARD separately. We propose a model in which filament formation is the first level of organization in the ASC speck. Filaments further compact in a higher organization level, which can be inhibited by the … dinkydoo fabrics and notionsWebFeb 25, 2024 · ASC oligomerization is a critical and unifying step in inflammasome activation [ 2 ]. Therefore, assays that measure oligomerization of ASC have been widely used to study inflammasome activation. These include, among others, measurements of ASC speck formation by microscopy, ASC oligomerization by Western blotting or flow … dinky donkey shropshireWebJul 15, 2013 · The ASC speck formation assay also supported this finding. ASC assembles into a large protein complex, or "speck", that is considered to be an upstream readout for … dinky doodle in the huntWebApr 4, 2024 · Speck assay and analysis ASC-Cerulean expressing THP1 cells were fixed with 4% formaldehyde for 15 min at room temperature, washed with PBS and stained with HCS Red (Invitrogen, Loughborough,... fortnite season chapter 2 season 3WebSep 4, 2024 · Speck formation in vivo is induced by NLR or ASC overexpression When ASC is present at endogenous concentrations, activated members of the NLR protein family, … dinky doughnuts cereal