Anti-IRAK (CT) Antibody (2426)
$445.00
SKU: 2426
Categories: Antibody Products, Neuroscience and Signal Transduction Antibodies, Products
Overview
Product Name Anti-IRAK (CT) Antibody (2426)
Description Anti-IRAK (CT) Rabbit Polyclonal Antibody
Target IRAK (CT)
Species Reactivity Human
Applications ELISA,WB,ICC,IP,IF
Host Rabbit
Clonality Polyclonal
Isotype IgG
Immunogen Peptide corresponding the C- terminus of human IRAK.
Properties
Form Liquid
Concentration Lot Specific
Formulation PBS, pH 7.4.
Buffer Formulation Phosphate Buffered Saline
Buffer pH pH 7.4
Format Purified
Purification Purified by peptide immuno-affinity chromatography
Specificity Information
Specificity This antibody recognizes full-length human IRAK (80 kD).
Target Name Interleukin-1 receptor-associated kinase 1
Target ID IRAK (CT)
Uniprot ID P51617
Alternative Names IRAK-1, EC 2.7.11.1
Gene Name IRAK1
Gene ID 3654
Accession Number NP_001020413
Sequence Location Cytoplasm, Nucleus, Lipid droplet
Biological 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. {PubMed:11397809, PubMed:12860405, PubMed:14684752, PubMed:15084582, PubMed:15465816, PubMed:15767370, PubMed:17997719, PubMed:20400509}.
Research Areas Neuroscience
Background Nuclear factor kappa B (NF-kappaB) is a ubiquitous transcription factor and key mediator of gene expression during immune and inflammatory responses. NF-kappaB activates numerous genes in response to extracellular stimuli, such as IL-1, TNFalpha, LPS, oxidative stress, and mitogens. NF-kappaB is associated with IkappaB in cytoplasm. After stimulation, 1kappaB is phosphorylated and dissociates from NF-kappaB. NF-kappaB enters cell nuclei where it activates an array of genes. A serine/threonine protein kinase associated with IL-1 receptor (IRAK) mediates a signaling cascade leading to NF-kappaB activation. IRAK is associated with IL-1 receptor subunits IL-1RI and IL-1RacP and serves as a signaling molecule to mediate IL-1 responses. IRAK also mediates IL-18-induced NF-kappaB activation.
Application Images
Description Western Blot Validation in Human Cell Lines
Loading: 15 ug of lysates per lane. Antibodies: IRAK 2426 (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Loading: 15 ug of lysates per lane. Antibodies: IRAK 2426 (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Description Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines
Loading: 15 ug of lysates per lane. Antibodies: IRAK 2426 (1 ug/mL), IRAK 64-231 (2 ug/mL), beta-actin (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Loading: 15 ug of lysates per lane. Antibodies: IRAK 2426 (1 ug/mL), IRAK 64-231 (2 ug/mL), beta-actin (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Description Western Blot Validation with Recombinant Protein
Loading: 30 ng of human IRAK recombinant protein per lane. Antibodies: IRAK 2426 (1: 1 ug/mL, 2: 2 ug/mL and 3: 4 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Loading: 30 ng of human IRAK recombinant protein per lane. Antibodies: IRAK 2426 (1: 1 ug/mL, 2: 2 ug/mL and 3: 4 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Description Species Activity in Mouse and Rat Cell Lines
Loading: 15 ug of lysates per lane. Antibodies: IRAK 2426 (1 µg/mL,), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Loading: 15 ug of lysates per lane. Antibodies: IRAK 2426 (1 µg/mL,), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Description Immunofluorescence Validation of IRAK in Human HeLa Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa Cells labeling IRAK with 2426 at 20 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa Cells labeling IRAK with 2426 at 20 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
Description Immunocytochemistry Validation of IRAK in Human HeLa Cells
Immunocytochemical analysis of HeLa cells using anti-IRAK antibody (2426) at 10 ug/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.
Immunocytochemical analysis of HeLa cells using anti-IRAK antibody (2426) at 10 ug/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.
Description 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 (2426). Ubiquitination was specifically detected by IRAK-1 immunoprecipitation followed by Western blotting with anti-HA antibodies.
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 (2426). Ubiquitination was specifically detected by IRAK-1 immunoprecipitation followed by Western blotting with anti-HA antibodies.
Description 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 β-actin expression levels by immunoblotting. IRAK1 signal was disrupted in IRAK1 KD lysate.
Chondrocytes were transfected with 250 nM of IRAK1 or control siRNA for 48 h and lysates were analyzed for IRAK1 or β-actin expression levels by immunoblotting. IRAK1 signal was disrupted in IRAK1 KD lysate.
Handling
Storage This antibody is stable for at least one (1) year at -20°C. Avoid multiple freeze-thaw cycles.
Dilution Instructions Dilute in PBS or medium which is identical to that used in the assay system.
Application Instructions 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.
Immunoprecipitation: use 2-4 ug antibody per sample.
Positive control: Whole cell lysate from HeLa cells or THP-1 cells.
References & Data Sheet
Data Sheet Download PDF Data Sheet