IRAK Antibody | 1007

IRAK Antibody | 1007 | Gentaur UK, US & Europe Distribution

Host: Rabbit

Reactivity: Human, Mouse, Rat

Homology: Predicted species reactivity based on immunogen sequence: Bovine: (85%)

Immunogen: Anti-IRAK antibody (1007) 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.

Research Area: Signal Transduction, Innate Immunity

Tested Application: E, WB, ICC, IP, IF

Application: 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.

Specificiy: At least four isoforms of IRAK are known to exist; this antibody will detect all four isoforms. IRAK antibody is predicted to not cross-react with other members of the IRAK protein family.

Positive Control 1: Cat. No. 1201 - HeLa Cell Lysate

Positive Control 2: Cat. No. 17-001 - HeLa Cell Slide

Positive Control 3: Cat. No. 1220 - SK-N-SH Cell Lysate

Positive Control 4: N/A

Positive Control 5: N/A

Positive Control 6: N/A

Molecular Weight: Predicted: 77kD
Observed: 77 kD

Validation:

Independent Antibody Validation in Cell lines (Figure 2) shows similar IRAK expression profile in human cell lines detected by two independent anti-IRAK antibodies that recognize different epitopes, 1007 against C-terminus domain and 64-231 against another region of C-terminus domain.  IRAK proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies.

Recombinant Protein Test (Figure 3): Anti-IRAK antibodies (1007) detected human IRAK recombinant protein at different concentrations.

Immunoprecipitation validation (Figure 6): IRAK1 protein and pellino protein were immunoprecipitated by anti-IRAK antibodies (1007) in HEK293T cells with co-expression of Pellino proteins and IRAK-1.

Overexpression validation (Figure 7): IRAK1 overexpression in 293T cells was detected by anit-IRAK antibodies (1007).

KD validation (Figure 8): Anti-IRAK antibody (1007) specificity was further verified by IRAK specific knockdown. IRAK signal in human chondrocytes transfected with IRAK siRNAs was disrupted in comparison with that in cells transfected with control siRNAs.

Isoform: Human IRAK has 4 isoforms, including isoform 1 (712aa, 77kD), isoform 2 (682aa, 73kD), isoform 3 (693aa, 75kD), and isoform 4 (633aa, 68kD). Mouse IRAK has 2 isoforms, including isoform 1 (710aa, 77kD) and isoform 2 (712aa, 78kD). Rat IRAK also has one isoform (710aa, 78kD). 1007 can detect all human isoforms, mouse and rat IRAK.

Purification: IRAK Antibody is affinity chromatography purified via peptide column.

Clonality: Polyclonal

Clone: N/A

Isotype: IgG

Conjugate: Unconjugated

Physical State: Liquid

Buffer: IRAK Antibody is supplied in PBS containing 0.02% sodium azide.

Concentration: 1 mg/mL

Storage Condition: 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.

Alternate Name: IRAK Antibody: IRAK, pelle, IRAK, Interleukin-1 receptor-associated kinase 1, IRAK-1

User Note: Optimal dilutions for each application to be determined by the researcher.

BACKGROUND: IRAK Antibody: Nuclear factor kappa B (NF-κB) is a ubiquitous transcription factor and an essential mediator of gene expression during activation of immune and inflammatory responses. NF-κB mediates the expression of a great variety of genes in response to extracellular stimuli including IL-1, TNFα and LPS. A serine/threonine protein kinase associated with IL-1 receptor (IRAK) and its homologue mouse pelle-like protein kinase (mPLK) were identified recently. IRAK is associated with the IL-1 receptor subunits IL-1RI and IL-1RAcP after IL-1 binding and serves as a signaling molecule to mediate IL-1 response. IRAK mediates a signaling cascade leading to NF-κB activation by members in IL-1 family including IL-1 and a novel cytokine IL-18 (also termed IGIF).