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rabbit anti-ASC polyclonal antibody 1972

$445.00

Antibody summary

  • Rabbit polyclonal to ASC
  • Suitable for: ELISA,WB,ICC,IHC-P
  • Isotype: IgG
  • 100 µg
SKU: 1972parent Category: Tags: , ,
Weight1 lbs
Dimensions9 × 5 × 2 in
host

rabbit

isotype

IgG

clonality

polyclonal

concentration

1 mg/mL

applications

ICC/IF, WB

reactivity

ASC

available sizes

100 µg

rabbit anti-ASC polyclonal antibody 1972

antibody
Tested applications
WB,IHC,IHC,ICC/IF,ELISA
Recommended dilutions
Immunoblotting: use at 1ug/mL.

Immunocytochemistry: use at 5ug/mL.

These are recommended concentrations.

Enduser should determine optimal concentrations for their applications.

Positive control: Whole cell lysate of HL60 cells.
Immunogen
Peptide corresponding to aa 182-195 of human ASC.
Size and concentration
100µg and lot specific
Form
liquid
Storage Instructions
This antibody is stable for at least one (1) year at -20°C. Avoid multiple freeze-thaw cycles.
Storage buffer
PBS, pH 7.4.
Purity
peptide affinty purifcation
Clonality
polyclonal
Isotype
IgG
Compatible secondaries
goat anti-rabbit IgG, H&L chain specific, peroxidase conjugated, conjugated polyclonal antibody 9512
goat anti-rabbit IgG, H&L chain specific, biotin conjugated polyclonal antibody 2079
goat anti-rabbit IgG, H&L chain specific, FITC conjugated polyclonal antibody 7863
goat anti-rabbit IgG, H&L chain specific, Cross Absorbed polyclonal antibody 2371
goat anti-rabbit IgG, H&L chain specific, biotin conjugated polyclonal antibody, crossabsorbed 1715
goat anti-rabbit IgG, H&L chain specific, FITC conjugated polyclonal antibody, crossabsorbed 1720
Isotype control
Rabbit polyclonal - Isotype Control
target relevance
Protein names
Apoptosis-associated speck-like protein containing a CARD (hASC) (Caspase recruitment domain-containing protein 5) (PYD and CARD domain-containing protein) (Target of methylation-induced silencing 1)
Gene names
PYCARD,PYCARD ASC CARD5 TMS1
Mass
21627Da
Function
FUNCTION: Functions as a key mediator in apoptosis and inflammation (PubMed:17599095, PubMed:25847972, PubMed:19494289, PubMed:15030775, PubMed:17349957, PubMed:19158675, PubMed:19158676, PubMed:30674671, PubMed:34678144, PubMed:24630722, PubMed:21487011, PubMed:19234215, PubMed:11103777, PubMed:12646168). Promotes caspase-mediated apoptosis involving predominantly caspase-8 and also caspase-9 in a probable cell type-specific manner (PubMed:11103777, PubMed:12646168). Involved in activation of the mitochondrial apoptotic pathway, promotes caspase-8-dependent proteolytic maturation of BID independently of FADD in certain cell types and also mediates mitochondrial translocation of BAX and activates BAX-dependent apoptosis coupled to activation of caspase-9, -2 and -3 (PubMed:16964285, PubMed:14730312). Involved in innate immune response by acting as an integral adapter in the assembly of various inflammasomes (NLRP1, NLRP2, NLRP3, NLRP6, AIM2 and probably IFI16) which recruit and activate caspase-1 leading to processing and secretion of pro-inflammatory cytokines (PubMed:17599095, PubMed:25847972, PubMed:15030775, PubMed:17349957, PubMed:19158675, PubMed:19158676, PubMed:30674671, PubMed:34678144, PubMed:16982856, PubMed:24630722, PubMed:21487011, PubMed:19234215, PubMed:23530044, PubMed:29440442, PubMed:33980849). Caspase-1-dependent inflammation leads to macrophage pyroptosis, a form of cell death (PubMed:24630722). The function as activating adapter in different types of inflammasomes is mediated by the pyrin and CARD domains and their homotypic interactions (PubMed:19234215, PubMed:14499617, PubMed:24630722). Clustered PYCARD nucleates the formation of caspase-1 filaments through the interaction of their respective CARD domains, acting as a platform for of caspase-1 polymerization (PubMed:24630722). In the NLRP1 and NLRC4 inflammasomes seems not be required but facilitates the processing of procaspase-1 (PubMed:17349957). In cooperation with NOD2 involved in an inflammasome activated by bacterial muramyl dipeptide leading to caspase-1 activation (PubMed:16964285). May be involved in RIGI-triggered pro-inflammatory responses and inflammasome activation (PubMed:19915568). In collaboration with AIM2 which detects cytosolic double-stranded DNA may also be involved in a caspase-1-independent cell death that involves caspase-8 (PubMed:19158675, PubMed:19158676). In adaptive immunity may be involved in maturation of dendritic cells to stimulate T-cell immunity and in cytoskeletal rearrangements coupled to chemotaxis and antigen uptake may be involved in post-transcriptional regulation of the guanine nucleotide exchange factor DOCK2; the latter function is proposed to involve the nuclear form (PubMed:22732093). Also involved in transcriptional activation of cytokines and chemokines independent of the inflammasome; this function may involve AP-1, NF-kappa-B, MAPK and caspase-8 signaling pathways (PubMed:12486103, PubMed:16585594). For regulation of NF-kappa-B activating and inhibiting functions have been reported (PubMed:12486103). Modulates NF-kappa-B induction at the level of the IKK complex by inhibiting kinase activity of CHUK and IKBK (PubMed:12486103, PubMed:16585594). Proposed to compete with RIPK2 for association with CASP1 thereby down-regulating CASP1-mediated RIPK2-dependent NF-kappa-B activation and activating interleukin-1 beta processing (PubMed:16585594). Modulates host resistance to DNA virus infection, probably by inducing the cleavage of and inactivating CGAS in presence of cytoplasmic double-stranded DNA (PubMed:28314590). {ECO:0000269|PubMed:11103777, ECO:0000269|PubMed:12486103, ECO:0000269|PubMed:12646168, ECO:0000269|PubMed:14499617, ECO:0000269|PubMed:14730312, ECO:0000269|PubMed:15030775, ECO:0000269|PubMed:16585594, ECO:0000269|PubMed:16964285, ECO:0000269|PubMed:16982856, ECO:0000269|PubMed:17349957, ECO:0000269|PubMed:17599095, ECO:0000269|PubMed:19158675, ECO:0000269|PubMed:19158676, ECO:0000269|PubMed:19234215, ECO:0000269|PubMed:19494289, ECO:0000269|PubMed:19915568, ECO:0000269|PubMed:21487011, ECO:0000269|PubMed:22732093, ECO:0000269|PubMed:23530044, ECO:0000269|PubMed:24630722, ECO:0000269|PubMed:25847972, ECO:0000269|PubMed:28314590, ECO:0000269|PubMed:29440442, ECO:0000269|PubMed:30674671, ECO:0000269|PubMed:33980849, ECO:0000269|PubMed:34678144}.; FUNCTION: [Isoform 2]: May have a regulating effect on the function as inflammasome adapter. {ECO:0000269|PubMed:19759850, ECO:0000269|PubMed:20482797}.; FUNCTION: [Isoform 3]: Seems to inhibit inflammasome-mediated maturation of interleukin-1 beta. {ECO:0000269|PubMed:20482797}.
Subellular location
SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:11103777, ECO:0000269|PubMed:15030775, ECO:0000269|PubMed:19234215, ECO:0000269|PubMed:25847972}. Inflammasome {ECO:0000269|PubMed:12191486, ECO:0000269|PubMed:15030775, ECO:0000269|PubMed:30674671, ECO:0000269|PubMed:34678144}. Endoplasmic reticulum {ECO:0000269|PubMed:21124315}. Mitochondrion {ECO:0000269|PubMed:14730312, ECO:0000269|PubMed:21124315}. Nucleus {ECO:0000269|PubMed:19234215, ECO:0000269|PubMed:25847972}. Note=Upstream of caspase activation, a redistribution from the cytoplasm to the aggregates occurs. These appear as hollow, perinuclear spherical, ball-like structures (PubMed:11103777, PubMed:12191486, PubMed:15030775). Upon NLRP3 inflammasome activation redistributes to the perinuclear space localizing to endoplasmic reticulum and mitochondria (PubMed:12191486, PubMed:15030775). Localized primarily to the nucleus in resting monocytes/macrophages and rapidly redistributed to the cytoplasm upon pathogen infection (PubMed:19234215). Localized to large cytoplasmic aggregate appearing as a speck containing AIM2, PYCARD, CASP8 and bacterial DNA after infection with Francisella tularensis (By similarity). {ECO:0000250|UniProtKB:Q9EPB4, ECO:0000269|PubMed:11103777, ECO:0000269|PubMed:12191486, ECO:0000269|PubMed:15030775, ECO:0000269|PubMed:19234215}.; SUBCELLULAR LOCATION: Golgi apparatus membrane {ECO:0000269|PubMed:23229815}. Note=(Microbial infection) Upon HRSV infection, the protein is mainly located in lipid rafts in the Golgi membrane. {ECO:0000269|PubMed:23229815}.
Tissues
TISSUE SPECIFICITY: 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. Expressed in lung epithelial cells (at protein level) (PubMed:23229815). 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. {ECO:0000269|PubMed:23229815}.
Structure
SUBUNIT: Self-associates; enforced oligomerization induces apoptosis, NF-kappa-B regulation and interleukin-1 beta secretion (PubMed:15641782, PubMed:17599095, PubMed:33420028, PubMed:33420033). Homooligomers can form disk-like particles of approximately 12 nm diameter and approximately 1 nm height (PubMed:15641782, PubMed:17599095). Next to isoform 1, also isoform 2 and isoform 3 may be involved in oligomerization leading to functional regulation (Probable). Component of several inflammasomes containing one pattern recognition receptor/sensor, such as NLRP1, NLRP2, NLRP3, NLRP6, NLRC4, AIM2, MEFV or NOD2, and probably NLRC4, NLRP12 or IFI16 (PubMed:11374873, PubMed:12191486, PubMed:15030775, PubMed:15456791, PubMed:19158676, PubMed:23530044, PubMed:27432880, PubMed:29440442, PubMed:30674671, PubMed:34678144, PubMed:35559676, PubMed:33980849). Major component of the ASC pyroptosome, a 1-2 um supramolecular assembly (one per macrophage cell) which consists of oligomerized PYCARD dimers and CASP1 (PubMed:17599095). Interacts with CASP1 (precursor form); the interaction induces activation of CASP1 leading to the processing of interleukin-1 beta; PYCARD competes with RIPK2 for binding to CASP1 (PubMed:11967258, PubMed:14634131, PubMed:16585594, PubMed:17599095, PubMed:33420033). Interacts with NLRP3; the interaction requires the homooligomerization of NLRP3 (PubMed:11786556, PubMed:15030775, PubMed:15020601, PubMed:34341353, PubMed:35559676). Interacts with NLRP2, NLRC4, MEFV, CARD16, AIM2, IFI16, NOD2, RIGI, RIPK2, PYDC1, PYDC2, NLRP10, CASP8, CHUK, IKBKB and BAX (PubMed:11374873, PubMed:11498534, PubMed:12486103, PubMed:12646168, PubMed:12656673, PubMed:15456791, PubMed:14730312, PubMed:15096476, PubMed:17178784, PubMed:17339483, PubMed:18362139, PubMed:19158675, PubMed:19158676, PubMed:19915568, PubMed:21575908, PubMed:23530044, PubMed:29440442, PubMed:33980849). Component of the AIM2 PANoptosome complex, a multiprotein complex that drives inflammatory cell death (PANoptosis) (By similarity). {ECO:0000250|UniProtKB:Q9EPB4, ECO:0000269|PubMed:11374873, ECO:0000269|PubMed:11498534, ECO:0000269|PubMed:11786556, ECO:0000269|PubMed:11967258, ECO:0000269|PubMed:12191486, ECO:0000269|PubMed:12486103, ECO:0000269|PubMed:12646168, ECO:0000269|PubMed:12656673, ECO:0000269|PubMed:14634131, ECO:0000269|PubMed:14730312, ECO:0000269|PubMed:15020601, ECO:0000269|PubMed:15030775, ECO:0000269|PubMed:15096476, ECO:0000269|PubMed:15456791, ECO:0000269|PubMed:15641782, ECO:0000269|PubMed:16585594, ECO:0000269|PubMed:17178784, ECO:0000269|PubMed:17339483, ECO:0000269|PubMed:17599095, ECO:0000269|PubMed:18362139, ECO:0000269|PubMed:19158675, ECO:0000269|PubMed:19158676, ECO:0000269|PubMed:19915568, ECO:0000269|PubMed:21575908, ECO:0000269|PubMed:23530044, ECO:0000269|PubMed:27432880, ECO:0000269|PubMed:29440442, ECO:0000269|PubMed:30674671, ECO:0000269|PubMed:33420028, ECO:0000269|PubMed:33420033, ECO:0000269|PubMed:33980849, ECO:0000269|PubMed:34341353, ECO:0000269|PubMed:34678144, ECO:0000269|PubMed:35559676}.
Post-translational modification
PTM: Phosphorylated. {ECO:0000269|PubMed:12656673}.; PTM: 'Lys-63'-linked polyubiquitination by TRAF3 is critical for speck formation and inflammasome activation. {ECO:0000269|PubMed:25847972}.
Target Relevance information above includes information from UniProt accession : Q9ULZ3
The UniProt Consortium

Data

benchmark-antibodies_anti-asc_antibody_1972_1.gif
Western Blot Validation in Human HL60 Cells
Loading: 15 µg of lysates per lane. Antibodies: ASC 1972, (1 µg/mL) in the absence (A) or presence (B) of blocking peptide, 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-asc_antibody_1972_2.gif
Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines
Loading: 15 µg of lysates per lane. Antibodies: ASC 1972, (2 µg/mL), ASC 39-001, (2 µg/mL), beta-actin (1 µg/mL) and GAPDH (0.02 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit or goat anti-mouse (for ASC 39001) IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-asc_antibody_1972_3.gif
Western Blot Validation in Human THP-1 Cells
Loading: 15 µg of lysate per lane. Antibodies: ASC 1972, (2 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-asc_antibody_1972_4.gif
Immunohistochemistry Validation of ASC in Human Spleen Tissue
Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-ASC antibody (1972) at 2.5 µg/mL. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
benchmark-antibodies_anti-asc_antibody_1972_5.gif
Immunofluorescence Validation of ASC in Human Spleen Tissue
Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Spleen Tissue labeling ASC with 1972 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
benchmark-antibodies_anti-asc_antibody_1972_6.gif
Immunocytochemistry Validation of ASC in HL60 Cells
Immunocytochemical analysis of HL60 cells using anti-ASC antibody (1972) at 5 µg/mL. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
benchmark-antibodies_anti-asc_antibody_1972_7.gif
KD Validation of ASC in THP-1 Cells (Dowds et al., 2004)
Immunofluorescence analysis with anti-ASC antibodies (1972) was performed for BIM in 293 cells transfected with GFP siRNA or ASC siRNA. ASC expression was disrupted after ASC siRNA knockdown.
benchmark-antibodies_anti-asc_antibody_1972_8.gif
Overexpression Validation of ASC in HEK293T Cells (Dowds et al., 2004)
Western blot analysis with anti-ASC antibodies (1972) was performed for ASC in HEK293T cells transfected with pcDNA3-ASC.

Publications

Published literature highly relevant to the biological target of this product and referencing this antibody or clone are retrieved from PubMed database provided by The United States National Library of Medicine at the National Institutes of Health.




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Protocols

relevant to this product
Western blot
IHC
ICC

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