Anti-IRAK-4 Antibody (56412)
$445.00
| Host | Quantity | Applications | Species Reactivity | Data Sheet | |
|---|---|---|---|---|---|
| Rabbit | 100ug | ELISA,WB,ICC,IF | Human |  |
Overview
Product Name Anti-IRAK-4 Antibody (56412)
Description Anti-IRAK-4 Rabbit Polyclonal Antibody
Target IRAK-4
Species Reactivity Human
Applications ELISA,WB,ICC,IF
Host Rabbit
Clonality Polyclonal
Isotype IgG
Immunogen A synthetic peptide corresponding to amino acids at the carboxy terminus of human IRAK-4.
Properties
Form Liquid
Concentration 1.0 mg/mL
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 specifically recognizes human IRAK-4 (IL-1 Receptor Associated Kinase-4). This antibody does not cross-react with other IRAKs.
Target Name Interleukin-1 receptor-associated kinase 4
Target ID IRAK-4
Uniprot ID Q9NWZ3
Alternative Names IRAK-4, EC 2.7.11.1, Renal carcinoma antigen NY-REN-64
Gene Name IRAK4
Sequence Location Cytoplasm
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 (PubMed:17878374). Is rapidly recruited by MYD88 to the receptor-signaling complex upon TLR activation to form the Myddosome together with IRAK2. Phosphorylates initially IRAK1, thus stimulating the kinase activity and intensive autophosphorylation of IRAK1. 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 NCF1 and regulates NADPH oxidase activation after LPS stimulation suggesting a similar mechanism during microbial infections. {PubMed:11960013, PubMed:12538665, PubMed:15084582, PubMed:17217339, PubMed:17337443, PubMed:17878374, PubMed:17997719, PubMed:20400509, PubMed:24316379}.
Research Areas Neuroscience
Background IRAK-4 activates the NF-ï«B and MAPK pathways and plays a role in IL-1R mediated inflammatory responses and innate immunity. IRAK-4-deficient animals are resistant to challenge with LPS and are impaired in their responses to viral and bacterial challenges.
Application Images
Description Western Blot Validation in Human Cell Lines
Loading: 15 ug/ of lysates per lane. Antibodies: IRAK4 56412 (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: IRAK4 56412 (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: IRAK4-56412 (1 ug/mL), IRAK4-24-025 (1 ug/mL), beta-actin (1.5 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: IRAK4-56412 (1 ug/mL), IRAK4-24-025 (1 ug/mL), beta-actin (1.5 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 in Human HeLa Cell Lines
Loading: 15 ug of lysates per lane. Antibodies: IRAK4 56412, 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Lane 1: 1 ug/mLLane 2: 2 ug/mLLane 3: 4 ug/mL
Loading: 15 ug of lysates per lane. Antibodies: IRAK4 56412, 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Lane 1: 1 ug/mLLane 2: 2 ug/mLLane 3: 4 ug/mL
Description Immunocytochemistry Validation of IRAK4 in K562 Cells
Immunocytochemical analysis of K562 cells using anti-IRAK4 antibody (56412) 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 K562 cells using anti-IRAK4 antibody (56412) 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 Immunofluorescence Validation of IRAK4 in K562 Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 Cells labeling IRAK4 with 56412 at 10 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 Cells labeling IRAK4 with 56412 at 10 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
Description KO and KD Validation of IRAK4 in Mouse Bone Marrow-derived Macrophages (BMDM) (Koziczak-Holbro et al., 2008)
Western blot analysis with anti-IRAK4 antibodies was performed for IRAK4 in BMDM isolated from the mice. IRAK4 expression was not observed in the IRAK-4-/- cells and also reduced in IRAK4 mutant (IRAK-4 KD) when compared with WT.
Western blot analysis with anti-IRAK4 antibodies was performed for IRAK4 in BMDM isolated from the mice. IRAK4 expression was not observed in the IRAK-4-/- cells and also reduced in IRAK4 mutant (IRAK-4 KD) when compared with WT.
Description Regulated Expression Validation of IRAK4 in Mouse RAW 264 Cells (Hatao et al., 2004)
RAW 264 cells were stimulated with (a) CSK4 and (b) CpG-DNA in the absence or presence of proteasome inhibitors (MG132 and Lactactstin). When detected with anti-IRAK4 antibodies (C12 from , Inc.), IRAK4 expression was found to be reduced without the inhibitors. The smaller protein band, a cleavage product of IRAK4, was present in the absence of both inhibitors.
RAW 264 cells were stimulated with (a) CSK4 and (b) CpG-DNA in the absence or presence of proteasome inhibitors (MG132 and Lactactstin). When detected with anti-IRAK4 antibodies (C12 from , Inc.), IRAK4 expression was found to be reduced without the inhibitors. The smaller protein band, a cleavage product of IRAK4, was present in the absence of both inhibitors.
Description KD Validation of IRAK4 in HEK293T Cells (Heinz et al., 2012)
Western blot analysis with anti-IRAK4 antibodies was performed for IRAK4 in HEK293T cells. IRAK4 expression was not observed in IRAK4 knockdown cells.
Western blot analysis with anti-IRAK4 antibodies was performed for IRAK4 in HEK293T cells. IRAK4 expression was not observed in IRAK4 knockdown cells.
Handling
Storage Stable for one year at -20°C. Store in appropriate aliquots to avoid multiple freeze-thaw cycles.
Dilution Instructions Dilute in PBS or medium that is identical to that used in the assay system.
Application Instructions Immunoblotting: Use at 1-4ug/mL. A band of approximately 50kDa is detected.
Immunocytochemistry: Use at 10ug/mL.
Immunofluorescence: Use at 10ug/mL.
These are recommended concentrations.
Enduser should determine optimal concentrations for their applications.
Immunocytochemistry: Use at 10ug/mL.
Immunofluorescence: Use at 10ug/mL.
These are recommended concentrations.
Enduser should determine optimal concentrations for their applications.
References & Data Sheet
Data Sheet 
Download PDF Data Sheet
Download PDF Data Sheet