IRAK1 Antibody

• Catalog Number: orb1239517
• Category: Antibodies
• Description: IRAK1 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, ICC, IF, IP, WB
• Predicted Reactivity: Bovine
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: Anti-IRAK antibody (orb1239517) was raised against a peptide corresponding to 13 amino acids near the carboxy terminus of human IRAK. The immunogen is located within the last 50 amino acids of IRAK.
• Concentration: 1 mg/mL
• Dilution range: WB: 1-4 μg/mL; IF: 20 μg/mL; ICC: 10 μg/mL.Antibody validated: Western Blot in human, mouse and rat samples; Immunofluorescence and Immunocytochemistry in human samples. All other applications and species not yet tested.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 77kDObserved: 77 kD
• Target: IRAK1
• UniProt ID: P51617
• NCBI: P51617
• Storage: IRAK antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
• Buffer/Preservatives: IRAK Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: IRAK Antibody: IRAK, pelle, IRAK, Interleukin-1 re
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Western Blot Validation in Human Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: IRAK orb1239517 (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: IRAK orb1239517 (1 µg/mL), IRAK orb1271054 (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.

Western Blot Validation with Recombinant Protein. Loading: 30 ng of human IRAK recombinant protein per lane. Antibodies: IRAK orb1239517 (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.

Species Activity in Mouse and Rat Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: IRAK orb1239517 (1 µg/mL, ), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Immunofluorescence Validation of IRAK in Human HeLa Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa Cells labeling IRAK with orb1239517 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

Immunocytochemistry Validation of IRAK in Human HeLa Cells. Immunocytochemical analysis of HeLa cells using anti-IRAK antibody (orb1239517) 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.

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 (orb1239517). Ubiquitination was specifically detected by IRAK-1 immunoprecipitation followed by Western blotting with anti-HA antibodies.

KD Validation in Human Chondrocytes (Ahmad et al., 2007). 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.