IRAK4 Antibody

• Catalog Number: orb1239512
• Category: Antibodies
• Description: IRAK4 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, ICC, IF, WB
• Predicted Reactivity: Bovine, Mouse, Rat
• Reactivity: Human
• Isotype: IgG
• Immunogen: Anti-IRAK4 antibody (orb1239512) was raised against a peptide corresponding to 13 amino acids near the carboxy terminus of human IRAK4. The immunogen is located within the last 50 amino acids of IRAK4.
• Concentration: 1 mg/mL
• Dilution range: WB: 1-4 μg/mL; ICC: 10 μg/mL; IF: 10 μg/mL.Antibody validated: Western Blot in human samples; Immunocytochemistry in human samples and Immunofluorescence in human samples. All other applications and species not yet tested.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 52kDObserved: 55 kD
• Target: IRAK4
• UniProt ID: Q9NWZ3
• NCBI: AAM15772
• Storage: IRAK-4 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-4 Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: IRAK-4 Antibody: IPD1, REN64, IRAK-4, NY-REN-64, I
• 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: IRAK4 orb1239512 (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: IRAK4 orb1239512 (1 µg/mL), IRAK4 orb1255476 (1 µg/mL), beta-actin (1.5 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Western Blot Validation in Human HeLa Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: IRAK4 orb1239512, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 1 µg/mL, Lane 2: 2 µg/mL, Lane 3: 4 µg/mL.

Immunocytochemistry Validation of IRAK4 in K562 Cells. Immunocytochemical analysis of K562 cells using anti-IRAK4 antibody (orb1239512) 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.

Immunofluorescence Validation of IRAK4 in K562 Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 Cells labeling IRAK4 with orb1239512 at 10 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

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.

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), 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.

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.