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Anti asc

Manufactured by Merck Group
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Anti-ASC is a laboratory equipment product designed for researchers and scientists. It is used to detect and quantify the presence of ASC (Apoptosis-associated Speck-like Protein Containing a CARD) in biological samples. Anti-ASC provides a tool for investigating cellular signaling pathways and immune responses.

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Lab products found in correlation

Anti nalp3, by Cell Signaling Technology (1 mentions) Anti caspase 1, by Cell Signaling Technology (1 mentions) Il 1β, by Cell Signaling Technology (1 mentions) Anti il 1β, by Cell Signaling Technology (1 mentions) Il 1β, by R&D Systems (1 mentions) Tnf α, by R&D Systems (1 mentions) Interleukin 6 (il 6), by R&D Systems (1 mentions) Il 10, by R&D Systems (1 mentions) N storm system, by Nikon (1 mentions) Anti tubulin, by Thermo Fisher Scientific (1 mentions) Du 897 emccd camera, by Agilent Technologies (1 mentions) Sp8 confocal microscopy, by Leica (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Dapi blue, by Merck Group (1 mentions) Black 96 well plate, by Greiner (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Triton x 100, by Merck Group (1 mentions) Odyssey imaging system, by LI COR (1 mentions)

Close spelling variants of this product

Anti-integrin β4 (clone ASC-8; (2 citations) Mouse anti-Asc antibody (2 citations)

5 protocols using anti asc

1

Cytokine and Inflammasome Analysis

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Culture supernatants IL-1β, IL-6, IL-10, TNF-α levels were measured by IL-1β (R&D Systems, Minnneapolis, MN, USA, DY201), IL-6 (R&D Systems, DY206), IL-10 (R&D Systems, DY217B), TNF-α (R&D Systems, DY210) ELISA kit. Caspase-1, ASC, Nalp3 and IL-1β was detected by western blot with anti-caspase-1 (Cell Signaling Technology, 2225S), anti-ASC (Millipore, Billerica, MA, USA, AB3607), anti-Nalp3 (Cell Signaling Technology, 13158S), and anti-IL-1β.
Xing Y., Cao R, & Hu H.M. (2016). TLR and NLRP3 inflammasome-dependent innate immune responses to tumor-derived autophagosomes (DRibbles). Cell Death & Disease, 7(8), e2322-.
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2

NLRP3 Inflammasome Activation and Imaging

Cited 1 time
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BMDMs were seeded 24 h before stimulation. Cells were primed with LPS for 4 h, then nigericin was added to activate the NLRP3 inflammasome, and the cells were incubated at 37 °C for 15–45 min. Cells were fixed with 4% paraformaldehyde and processed for STORM imaging. The antibodies used for labelling were anti-DDX3 (Bethyl Laboratories, A300–474A), anti-ASC (Millipore, 04–147) and anti-tubulin (Thermo Fisher Scientific, clone YOL1/34; diluted 1:1,000). STORM acquisition of 10 fields per condition was performed using a Nikon N-STORM system consisting of an inverted TiE stand, a 100 X, 1.45 NA objective, a DU-897 EMCCD camera, an Agilent laser launch and appropriate optics, as previously reported42 (link). Images of single-molecule reconstructions were exported for downstream analysis by Imaris software. Coordinate (x, y) positions of individual molecules were exported for analysis with a new statistical measure for analysing colocalization in super-resolution images (r-index; X. Liu et al., unpublished protocol). With this method, a value of 1 indicates colocalization, a value of 0 indicates complete randomness and a value of − 1 indicates exclusion.
Samir P., Kesavardhana S., Patmore D.M., Gingras S., Malireddi R.K., Karki R., Guy C.S., Briard B., Place D.E., Bhattacharya A., Sharma B.R., Nourse A., King S.V., Pitre A., Burton A.R., Pelletier S., Gilbertson R.J, & Kanneganti T.D. (2019). DDX3X acts as a live-or-die checkpoint in stressed cells by regulating NLRP3 inflammasome. Nature, 573(7775), 590-594.
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3

Immunofluorescence Assay of Pyroptosis Proteins

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Primary BMDMs fully differentiated from wild-type mice were infected with S. Typhimurium for 1.5 h at a multiplicity of infection of 1, fixed in 4% paraformaldehyde, and permeabilized in 0.5% Triton X-100 for 10 min. Samples were then blocked for 1 h in 5% bovine serum albumin in PBS. Samples were incubated with the following primary antibodies overnight at 4°C: anti-ASC (Millipore 04-147, diluted 1:100), anti-cleaved CASP3 (CST, #9661, diluted 1:250), and anti-phospho-MLKL (Santa Cruz, sc-165025, diluted 1:250). Samples were incubated with the following secondary antibodies for 1 h at room temperature: Alexa Fluor 488-conjugated antibody against mouse immunoglobulin G (Invitrogen, A21202, diluted 1:250), Alexa Fluor 568-conjugated antibody against rabbit immunoglobulin G (Invitrogen, A10042, diluted 1:250), and Alexa Fluor 647-conjugated antibody against goat immunoglobulin G (Invitrogen, A21447, diluted 1:250). Cells were counterstained with DAPI (Life Technologies), and confocal microscopy was performed using the Leica SP-8 confocal microscopy. Images were analyzed using FIJI software (ImageJ).
Christgen S., Zheng M., Kesavardhana S., Karki R., Malireddi R.K., Banoth B., Place D.E., Briard B., Sharma B.R., Tuladhar S., Samir P., Burton A, & Kanneganti T.D. (2020). Identification of the PANoptosome: A Molecular Platform Triggering Pyroptosis, Apoptosis, and Necroptosis (PANoptosis). Frontiers in Cellular and Infection Microbiology, 10, 237.
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4

Fluorescence Imaging of T. cruzi Infection

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Cells were plated for 18 h in a 96-well black plate (Greiner) with a clear bottom for microscopy. On the next day cells were infected with T. cruzi as described above and after 4 h the supernatant was removed, and cells were fixed with 4% paraformaldehyde (Sigma Aldrich) diluted in PBS for at least 15 min. Then, wells were washed twice with warm PBS, followed by block/permeabilization buffer [10% BSA (Sigma Aldrich), 1% FBS (LGC), 0.5% Triton-X 100 (Sigma Aldrich), diluted in PBS] for 1 h at room temperature. Wells were carefully washed twice with warm PBS and incubated for 18 h at 4°C with 1:1000 anti-ASC (Millipore, clone 2EI-7). On the next day wells were washed again with warm PBS and incubated with secondary antibody Alexa-fluor 647 (red) (Invitrogen) 1:1000 for 1 h at room temperature. Wells were washed again, incubated with 5 μg/mL DAPI (blue) (Sigma Aldrich) and images were acquired on IN Cell Analyzer 2200 equipment.
Amaral M.P., Cardoso F.D., de Farias I.S., de Souza R.Q., Matteucci K.C., Torrecilhas A.C, & Bortoluci K.R. (2023). NAIP/NLRC4 inflammasome participates in macrophage responses to Trypanosoma cruzi by a mechanism that relies on cathepsin-dependent caspase-1 cleavage. Frontiers in Immunology, 14, 1282856.
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5

Western Blot Analysis of Immune Signaling

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Whole cell extracts were electrophoresed in 10% SDS-PAGE gels and transferred onto Immobilon-FL membranes (Millipore). Blots were incubated with antibodies specific for pTBK1 (Ser172), pIRF3 (Ser396), total TBK1, total IRF3, cGAS, STING, GAPDH or β-actin (Cell Signaling). For ASC-myc detection in ASC−/− macrophages, anti-ASC (Millipore) was used. For p204-myc detection in WT macrophages, anti-myc (Cell Signaling) was used. Anti-rabbit or anti-mouse IRDye 680RD label secondary antibodies were used for visualization of bands with the Odyssey Imaging system (LI-COR).
Corrales L., Woo S.R., Williams J., McWhirter S.M., Dubensky T.W, & Gajewski T.F. (2016). Antagonism of the STING pathway via activation of the AIM2 inflammasome by intracellular DNA. Journal of immunology (Baltimore, Md. : 1950), 196(7), 3191-3198.
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