Rabbit Asc Polyclonal Antibody | anti-Asc antibody

Asc (Tyr-144), phospho-specific Antibody

Cat. Num. AAA474195

• Gene Names: Pycard; Asc; TNS1; masc; CARD5; TMS-1; 9130417A21Rik
• Reactivity: Human, Rat, Mouse
• Applications: Western Blot, ELISA, Immunocytochemistry
• Synonyms: Asc; Polyclonal Antibody; Asc (Tyr-144); phospho-specific Antibody; phospho-specific; anti-Asc antibody

• Host: Rabbit
• Reactivity: Human, Rat, Mouse
• Clonality: Polyclonal
• Specificity: 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 20kDa Asc protein in western blots of J774A.1 mouse macrophages treated with pervanadate.
• Form/Format: Rabbit polyclonal, affinity-purified antibody is supplied in 100ul phosphate-buffered saline, 50% glycerol, 1mg/ml BSA, and 0.05% sodium azide.

• Applicable Applications for anti-Asc antibody: Western Blot (WB), ELISA (EIA), Immunocytochemistry (ICC)
• Application Notes: ELISA: 1:1000; ICC: 1:100; WB: 1:1000
• Immunogen: 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.
• Preparation and Storage: Store at -20 degree C. Stable for 1 year.

Western Blot (WB)

(Western blot analysis of mouse macrophage J774A.1 cells stimulated with pervanadate (1mM 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.)

Immunocytochemistry (ICC)

(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))

Related Product Information for anti-Asc antibody

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.

NCBI and Uniprot Product Information

• NCBI GI #: 165377185
• NCBI GeneID: 66824
• NCBI Accession #: NP_075747.3
• NCBI GenBank Nucleotide #: NM_023258.4
• UniProt Accession #: Q9EPB4; Q9D2W9
• Molecular Weight: 21,459 Da
• NCBI Official Full Name: apoptosis-associated speck-like protein containing a CARD
• NCBI Official Synonym Full Names: PYD and CARD domain containing
• NCBI Official Symbol: Pycard
• NCBI Official Synonym Symbols: Asc; TNS1; masc; CARD5; TMS-1; 9130417A21Rik
• NCBI Protein Information: apoptosis-associated speck-like protein containing a CARD
• UniProt Protein Name: Apoptosis-associated speck-like protein containing a CARD
• Protein Family: Ascalin
• UniProt Gene Name: Pycard
• UniProt Synonym Gene Names: Asc; mASC
• UniProt Entry Name: ASC_MOUSE

Uniprot Description

PYCARD: Promotes caspase-mediated apoptosis. This proapoptotic activity is mediated predominantly through the activation of caspase-9. May be a component of the inflammasome, a protein complex which also includes NALP2, CARD8 and CASP1 and whose function would be the activation of proinflammatory caspases. Forms complexes with other DAPIN domain-containing proteins. Interacts with CIAS1/PYPAF1 and PYDC1. Widely expressed at low levels. Detected in peripheral blood leukocytes, lung, small intestine, spleen, thymus, colon and at lower levels in placenta, liver and kidney. Very low expression in skeletal muscle, heart and brain. Detected in the leukemia cell lines HL-60 and U-937, but not in Jurkat T- cell lymphoma and Daudi Burkitt's lymphoma. Detected in the melanoma cell line WM35, but not in WM793. Not detected in HeLa cervical carcinoma cells and MOLT-4 lymphocytic leukemia cells. 3 isoforms of the human protein are produced by alternative splicing.

Protein type: Tumor suppressor; Adaptor/scaffold

Cellular Component: protein complex; mitochondrion; cell soma; endoplasmic reticulum; cytoplasm; extracellular region; nucleolus; IkappaB kinase complex; cytosol; nucleus

Molecular Function: protein binding; protein homodimerization activity; protease binding; apoptotic protease activator activity; Pyrin domain binding

Biological Process: myeloid dendritic cell activation during immune response; apoptosis; positive regulation of apoptosis; regulation of protein stability; positive regulation of caspase activity; positive regulation of JNK cascade; positive regulation of interleukin-1 beta secretion; defense response to Gram-negative bacterium; regulation of caspase activity; positive regulation of activated T cell proliferation; activation of NF-kappaB transcription factor; regulation of apoptosis; tumor necrosis factor-mediated signaling pathway; positive regulation of phagocytosis; inflammatory response; DNA damage response, signal transduction by p53 class mediator resulting in induction of apoptosis; defense response to virus; positive regulation of adaptive immune response; caspase activation; regulation of GTPase activity; immune system process; positive regulation of interleukin-6 production; interleukin-1 beta production; negative regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of tumor necrosis factor production; negative regulation of interferon-beta production; myeloid dendritic cell activation; positive regulation of interferon-gamma production; positive regulation of actin filament polymerization; response to bacterium; inhibition of NF-kappaB transcription factor; regulation of inflammatory response; innate immune response; positive regulation of transcription factor activity; regulation of autophagy; positive regulation of T cell activation; positive regulation of defense response to virus by host; activation of innate immune response; positive regulation of antigen processing and presentation of peptide antigen via MHC class II