IRAK4
Reactivity: Human, Mouse, Cow, Dog
WB, ELISA, IHC, IP, DB
Host: Mouse
Monoclonal
A8A8
unconjugated
Application Notes
This antibody conjugate is suitable for intracellular staining of human peripheral blood mononuclear cells and cell lines (using BD™ Phosflow Lyse/Fix Buffer or BD Cytofix™ Fixation Buffer), but not human whole blood. BD™ Phosflow Perm Buffer II or Perm Buffer III may be used.
Sample Volume
20 μL
Restrictions
For Research Use only
Format
Liquid
Buffer
Aqueous buffered solution containing BSA and ≤0.09 % sodium azide.
Preservative
Sodium azide
Precaution of Use
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
4 °C
Storage Comment
The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. Confirmation of the specificity of mAb L29-525 by western blot analysis (left panel). Lysates of human PBMC were probed with unconjugated antibody at a concentration of 0.1 μg/ml. IRAK4 is identified as a band of about 52 kDa. Western blot validation of IRAK4 by RNAi (right panel). Lysates from HeLa S3 cells (lane 1) and IRAK4 RNAi-transfected HeLa S3 cells (lane 2) were probed with unconjugated mAb L29-525 at 0.015 μg/ml. Down-regulation of IRAK4 expression is evident in the RNAi-transfected cells.
Suzuki, Saito: "IRAK-4--a shared NF-kappaB activator in innate and acquired immunity." in: Trends in immunology, Vol. 27, Issue 12, pp. 566-72, (2006) (PubMed).
Hatao, Muroi, Hiki, Ogawa, Mimura, Kaminishi, Tanamoto: "Prolonged Toll-like receptor stimulation leads to down-regulation of IRAK-4 protein." in: Journal of leukocyte biology, Vol. 76, Issue 4, pp. 904-8, (2004) (PubMed).
Lye, Mirtsos, Suzuki, Suzuki, Yeh: "The role of interleukin 1 receptor-associated kinase-4 (IRAK-4) kinase activity in IRAK-4-mediated signaling." in: The Journal of biological chemistry, Vol. 279, Issue 39, pp. 40653-8, (2004) (PubMed).
Li, Strelow, Fontana, Wesche: "IRAK-4: a novel member of the IRAK family with the properties of an IRAK-kinase." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 99, Issue 8, pp. 5567-72, (2002) (PubMed).
Interleukin 1 (IL-1) is a proinflammatory cytokine that mediates the host inflammatory response. IL-1 exerts its biological effects through its type I receptor (IL-1RI), which is found on most cell types. When IL-1 binds to the IL-1RI, NF-kappaB (transcription factor) is activated and translocates to the nucleus where it induces gene expression. IL-1-receptor-associated kinase (IRAK) has been reported to co-immunoprecipitate with IL-1RI. The IRAK family is made up of four identified Pelle homologs that regulate the activation of NF-kappaB and mitogen-activated protein (MAP) kinase pathways. IRAK4 is a unique member of the IRAK family because it is the closest mammalian homolog to Pelle. Also, when overexpressed, its kinase activity allows it to activate both the NF-kappaB and mitogen-activated protein (MAP) kinase pathways, and block the IL-1-induced activation and modification of IRAK1. IRAK4 is also able to phosphorylate IRAK1, which suggests that IRAK4 affects the early signal transduction of Toll/IL-1 receptors before the involvement of IRAK1. The L29-525 monoclonal antibody recognizes human IRAK4, regardless of phosphorylation status. The specificity of this antibody conjugate for flow cytometric analysis was validated by confirming that RNA-mediated interference (RNAi) of the specific protein reduced the staining of the cells (see figure). Furthermore, the capacity of the RNAi to down-regulate the expression of the relevant protein was confirmed by western blot analysis. Synonyms: IRAK-4, REN64, Q69FE3