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rabbit anti-IRAK (CT) polyclonal antibody 5960

$445.00

Antibody summary

  • Rabbit polyclonal to IRAK (CT)
  • Suitable for: ELISA,WB,ICC,IP,IF
  • Isotype: IgG
  • 100 µg
SKU: 5960parent Category: Tags: , ,
Weight1 lbs
Dimensions9 × 5 × 2 in
host

rabbit

isotype

IgG

clonality

polyclonal

concentration

1 mg/mL

applications

ICC/IF, WB

reactivity

IRAK (CT)

available sizes

100 µg

rabbit anti-IRAK (CT) polyclonal antibody 5960

antibody
Tested applications
WB,IHC,IHC,ICC/IF,ELISA,IP
Recommended dilutions
Immunoblotting: use at 1:1,000-1:2,000 dilution.

Immunoprecipitation: use 2-4 ug antibody per sample.

Positive control: Whole cell lysate from HeLa cells or THP-1 cells.
Immunogen
Peptide corresponding the C- terminus of human IRAK.
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
Interleukin-1 receptor-associated kinase 1 (IRAK-1) (EC 2.7.11.1)
Gene names
IRAK1,IRAK1 IRAK
Protein family
Protein kinase superfamily, TKL Ser/Thr protein kinase family, Pelle subfamily
Mass
76537Da
Function
FUNCTION: Serine/threonine-protein kinase that plays a critical role in initiating innate immune response against foreign pathogens. Involved in Toll-like receptor (TLR) and IL-1R signaling pathways. Is rapidly recruited by MYD88 to the receptor-signaling complex upon TLR activation. Association with MYD88 leads to IRAK1 phosphorylation by IRAK4 and subsequent autophosphorylation and kinase activation. Phosphorylates E3 ubiquitin ligases Pellino proteins (PELI1, PELI2 and PELI3) to promote pellino-mediated polyubiquitination of IRAK1. Then, the ubiquitin-binding domain of IKBKG/NEMO binds to polyubiquitinated IRAK1 bringing together the IRAK1-MAP3K7/TAK1-TRAF6 complex and the NEMO-IKKA-IKKB complex. In turn, MAP3K7/TAK1 activates IKKs (CHUK/IKKA and IKBKB/IKKB) leading to NF-kappa-B nuclear translocation and activation. Alternatively, phosphorylates TIRAP to promote its ubiquitination and subsequent degradation. Phosphorylates the interferon regulatory factor 7 (IRF7) to induce its activation and translocation to the nucleus, resulting in transcriptional activation of type I IFN genes, which drive the cell in an antiviral state. When sumoylated, translocates to the nucleus and phosphorylates STAT3. {ECO:0000269|PubMed:11397809, ECO:0000269|PubMed:12860405, ECO:0000269|PubMed:14684752, ECO:0000269|PubMed:15084582, ECO:0000269|PubMed:15465816, ECO:0000269|PubMed:15767370, ECO:0000269|PubMed:17997719, ECO:0000269|PubMed:20400509}.
Catalytic activity
CATALYTIC ACTIVITY: Reaction=ATP + L-seryl-[protein] = ADP + H(+) + O-phospho-L-seryl-[protein]; Xref=Rhea:RHEA:17989, Rhea:RHEA-COMP:9863, Rhea:RHEA-COMP:11604, ChEBI:CHEBI:15378, ChEBI:CHEBI:29999, ChEBI:CHEBI:30616, ChEBI:CHEBI:83421, ChEBI:CHEBI:456216; EC=2.7.11.1; CATALYTIC ACTIVITY: Reaction=ATP + L-threonyl-[protein] = ADP + H(+) + O-phospho-L-threonyl-[protein]; Xref=Rhea:RHEA:46608, Rhea:RHEA-COMP:11060, Rhea:RHEA-COMP:11605, ChEBI:CHEBI:15378, ChEBI:CHEBI:30013, ChEBI:CHEBI:30616, ChEBI:CHEBI:61977, ChEBI:CHEBI:456216; EC=2.7.11.1;
Subellular location
SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:16690127}. Nucleus {ECO:0000269|PubMed:16690127}. Lipid droplet {ECO:0000250}. Note=Translocates to the nucleus when sumoylated. RSAD2/viperin recruits it to the lipid droplet (By similarity). {ECO:0000250}.
Tissues
TISSUE SPECIFICITY: Isoform 1 and isoform 2 are ubiquitously expressed in all tissues examined, with isoform 1 being more strongly expressed than isoform 2. {ECO:0000269|PubMed:11397809}.
Structure
SUBUNIT: Homodimer (By similarity). Forms a complex with TRAF6, PELI1, IRAK4 and MYD88 (PubMed:16951688). Direct binding of SMAD6 to PELI1 prevents complex formation and hence negatively regulates IL1R-TLR signaling and eventually NF-kappa-B-mediated gene expression (PubMed:16951688). The TRAF6-PELI1-IRAK4-MYD88 complex recruits MAP3K7/TAK1, TAB1 and TAB2 to mediate NF-kappa-B activation (PubMed:16951688). Interaction with MYD88 recruits IRAK1 to the stimulated receptor complex (PubMed:9430229). Interacts with TOLLIP; this interaction occurs in the cytosol prior to receptor activation (PubMed:10854325). Interacts with IL1RL1 (PubMed:16286016). Interacts with PELI1 and TRAF6 (PubMed:12496252). Interacts (when polyubiquitinated) with IKBKG/NEMO (PubMed:18347055). Interacts with RSAD2/viperin (By similarity). Interacts with IRAK1BP1 (By similarity). Interacts with PELI2 (By similarity). Interacts with ZC3H12A; this interaction increases the interaction between ZC3H12A and IKBKB/IKKB (By similarity). Interacts with IRAK4 (PubMed:11960013). Interacts with PELI3 (PubMed:12874243). Interacts with INAVA; the interaction takes place upon PRR stimulation (PubMed:28436939). Interacts (via C-terminus) with NFATC4 (via N-terminus) (PubMed:18691762). {ECO:0000250|UniProtKB:Q62406, ECO:0000269|PubMed:10854325, ECO:0000269|PubMed:11960013, ECO:0000269|PubMed:12496252, ECO:0000269|PubMed:12874243, ECO:0000269|PubMed:16286016, ECO:0000269|PubMed:16951688, ECO:0000269|PubMed:18347055, ECO:0000269|PubMed:18691762, ECO:0000269|PubMed:28436939, ECO:0000269|PubMed:29883609, ECO:0000269|PubMed:9430229}.; SUBUNIT: (Microbial infection) Interacts with mumps virus protein SH; this interaction inhibits downstream NF-kappa-B pathway activation. {ECO:0000269|PubMed:28659487}.; SUBUNIT: (Microbial infection) Interacts with alphaviruses SINV, CHIKV, RRV, VEEV and EEEV capsid proteins; the interactions lead to inhibition of IRAK1-dependent signaling. {ECO:0000269|PubMed:33673546}.
Post-translational modification
PTM: Following recruitment on the activated receptor complex, phosphorylated on Thr-209, probably by IRAK4, resulting in a conformational change of the kinase domain, allowing further phosphorylations to take place. Thr-387 phosphorylation in the activation loop is required to achieve full enzymatic activity. {ECO:0000269|PubMed:11397809, ECO:0000269|PubMed:12538665, ECO:0000269|PubMed:14625308, ECO:0000269|PubMed:14684752}.; PTM: Polyubiquitinated by TRAF6 after cell stimulation with IL-1-beta by PELI1, PELI2 and PELI3. Polyubiquitination occurs with polyubiquitin chains linked through 'Lys-63'. Ubiquitination promotes interaction with NEMO/IKBKG. Also sumoylated; leading to nuclear translocation. {ECO:0000269|PubMed:16690127, ECO:0000269|PubMed:18347055, ECO:0000269|PubMed:19675569}.
Target Relevance information above includes information from UniProt accession : P51617
The UniProt Consortium

Data

benchmark-antibodies_anti-irak_ct_antibody_5960_1.gif
Western Blot Validation in Human Cell Lines
Loading: 15 µg of lysates per lane. Antibodies: IRAK 5960 (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-irak_ct_antibody_5960_2.gif
Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines
Loading: 15 µg of lysates per lane. Antibodies: IRAK 5960 (1 µg/mL), IRAK 64-231 (2 µg/mL), beta-actin (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-irak_ct_antibody_5960_3.gif
Western Blot Validation with Recombinant Protein
Loading: 30 ng of human IRAK recombinant protein per lane. Antibodies: IRAK 5960 (1: 1 µg/mL, 2: 2 µg/mL and 3: 4 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-irak_ct_antibody_5960_4.gif
Species Activity in Mouse and Rat Cell Lines
Loading: 15 µg of lysates per lane. Antibodies: IRAK 5960 (1 µg /mL,), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-irak_ct_antibody_5960_5.jpg
Immunofluorescence Validation of IRAK in Human HeLa Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa Cells labeling IRAK with 5960 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
benchmark-antibodies_anti-irak_ct_antibody_5960_6.jpg
Immunocytochemistry Validation of IRAK in Human HeLa Cells
Immunocytochemical analysis of HeLa cells using anti-IRAK antibody (5960) at 10 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 4 C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
benchmark-antibodies_anti-irak_ct_antibody_5960_7.gif
Immunoprecipitation and Overexpression Validation in HEK293T Cells(Schauvliege et al., 2006)
Co-expression of Pellino proteins and IRAK-1 leads to Pellino phosphorylation and IRAK-1 polyubiquitination. (A) E-tagged Pellino proteins were co-expressed with IRAK-1WT and HA?ubiquitin in HEK293T cells. For assessment of IRAK-1 polyubiquitination, the same cellextracts, untreated or treated with phosphatase as described above, were analysed for slower migrating forms of IRAK-1 by Western blotting withanti-IRAK-1 (5960). Ubiquitination was specifically detected by IRAK-1 immunoprecipitation followed by Western blotting with anti-HA antibodies.
benchmark-antibodies_anti-irak_ct_antibody_5960_8.gif
KD Validation in Human Chondrocytes (Ahmad et al., 2416)
Chondrocytes were transfected with 250 nM of IRAK1 or control siRNA for 48 h and lysates were analyzed for IRAK1 or ?-actin expression levels by immunoblotting. IRAK1 signal was disrupted in IRAK1 KD lysate.

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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