Catalog # AP5631

Asc (Tyr-144), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:1000
ICC	1:100
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 22 kDa

Host- and pathogen-associated cytoplasmic double-stranded DNA triggers the activation of a NALP3-independent inflammasome, which activates caspase-1, leading to maturation of pro-interleukin-1beta and inflammation. Several studies have isolated AIM2 (absent in melanoma 2) as a candidate cytoplasmic-DNA-sensing protein that contains an N-terminal pyrin domain and C-terminal oligonucleotide binding domain. A screen for transcripts induced by interferon-beta identified AIM2 gene expression. AIM2 protein bound double-stranded DNA, recruited the inflammasome adaptor ASC, and localized to ASC containing speckles. AIM2 and ASC form a pyroptosome, which induces pyroptotic cell death mediated by caspase-1. Asc can be phosphorylated at Tyr-144 in a Syk and JNK-dependent manner. This phosphorylation is critical for Asc speck formation and Caspase-1 activation.

References

Hara, H. et al. (2013) Nature Immunol. 14(12):1247.
Roberts, T.L. et al. (2009). Science. 323(5917):1057.
Bürckstümmer, T. et al. (2009). Nat Immunol. 10(3):266.

Western blot analysis of mouse macrophage J774A.1 cells stimulated with pervanadate (1 mM for 30 min.), then untreated (-) or treated (+) with alkaline phosphatase. The blot was probed with rabbit polyclonal anti-Asc (Tyr-144) phospho-specific antibody (AP5631) at 1:500.

Immunocytochemical labeling of Asc (Tyr-144) in inflammasomes. Paraformaldehyde fixed J774 cells were primed with LPS and treated with nigericin. Cells were co-labeled with DAPI, a caspase-1 inhibitor (FAM-YVAD-FMK), and anti-Asc (Tyr-144) phosphospecific antibody detected with AlexaFluor 568 secondary. (Image provided by Jordan Yaron, Center for Biosignatures Discovery Automation, Arizona State University)

Asc (Tyr-144) phospho-peptide (coupled to KLH) corresponding to amino acid residues surrounding Tyr-144 in mouse Asc. This peptide sequence is highly conserved in human and rat Asc.

*For more information, see UniProt Accession Q9EPB4

Rabbit polyclonal, affinity-purified antibody is supplied in 100 μl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

This antibody was cross-adsorbed to unphosphorylated Asc (Tyr-144) peptide before affinity purification using phospho-Asc (Tyr-144) peptide (without carrier). The antibody detects phosphorylated Asc (Tyr-144) in the inflammasome. In J774 macrophage cells primed with LPS and treated with nigericin, the antibody colocalized with an inflammasome marker, caspase-1 inhibitor (FAM-YVAD-FMK). In addition, the antibody detects a 20 kDa Asc protein in western blots of J774A.1 mouse macrophages treated with pervanadate.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

Yu, SH et al. (2019) J Ethnopharmacol. 239:111917. (WB: mouse bone marrow macrophages)
Dubois, EC et al. (2019) PLoS Pathog. 15(4):e1007709. (WB: mouse macrophage)
Furuya, MY et al. (2018) Arthritis Res Ther. 20(1):196. (WB: human neutrophils)
Kwak, SB et al. (2018) Mediators Inflamm. 2018:6054069. (WB: mouse J774A.1, BMDMs)
Hoyt, L.R. et al. (2016) J Immunol. 197:1322. (WB: mouse macrophage, LPS)

This kit contains: