Following infection, BMDCs were washed three times with PBS and incubated with media containing 1× FLICA far-red 660 active caspase-1 for 1 h (ImmunoChemistry Technologies). Cells were then fixed in 4% paraformaldehyde for 15 min, followed by blocking with 10% normal goat serum (Dako) in 0.1% saponin (Sigma) for 1 h. Cells were incubated with a mouse anti-Asc antibody (1:500 dilution, clone 2EI-7; Millipore) overnight followed by incubation with a rabbit anti-caspase-8 (1:500 dilution, 8592, CST) for an additional 1 h. The secondary antibodies used were Alexa Fluor 488 anti-rabbit IgG and Alexa Fluor 568 anti-mouse IgG. Cells were counterstained in DAPI mounting medium (1:250 dilution, Vecta Labs). Cells and inflammasomes were visualized, counted, and imaged using a Nikon C1 confocal microscope at the Cell and Tissue Imaging Center Light Microscopy Facility (CTIC-LM) at St. Jude.
Mouse anti asc antibody
Manufactured by Merck Group
The Mouse anti-Asc antibody is a laboratory reagent used for the detection and analysis of the Asc (Apoptosis-associated speck-like protein containing a CARD) protein. The antibody is designed to specifically bind to and identify the Asc protein, which is involved in the formation of the inflammasome complex and plays a role in the regulation of inflammatory responses.
Automatically generated - may contain errors
2 protocols using mouse anti asc antibody
1
Inflammasome Visualization and Quantification
2
Visualizing Inflammasome Formation in BMDMs
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Following infection, BMDMs were washed twice with PBS and incubated with media containing FAM FLICA caspase-1 for 1 h (ImmunoChemistry Technologies). Cells were then fixed in 4% paraformaldehyde for 15 min. Cells were incubated with a mouse anti-Asc antibody (1:500 dilution, clone 2EI-7; Millipore) overnight followed by incubation with a rabbit anti-caspase-8 (1:500 dilution, 8592; CST) for an additional 1 h. The secondary antibodies used were anti-rabbit Alexa Fluor 647 and anti-mouse Alexa Fluor 568. Cells were counterstained with DAPI. Cells and inflammasomes were visualized, counted, and imaged using a Nikon C2 confocal microscope at the Cell and Tissue Imaging Center Light Microscopy Facility (CTIC-LM) at St. Jude.
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