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Product Info Summary
| SKU: | A01021-2 |
|---|---|
| Size: | 0.1 mg |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, IP, IF, ICC, WB |
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Product info
Product Name
Anti-IRAK IRAK1 Antibody
SKU/Catalog Number
A01021-2
Size
0.1 mg
Form
Liquid
Description
Boster Bio Anti-IRAK IRAK1 Antibody (Catalog # A01021-2). Tested in ELISA, WB, ICC, IP, IF applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
IRAK antibody can be stored at 4°C for three months and -20°C, stable for up to one year. Avoid repeated freeze-thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Cite This Product
Anti-IRAK IRAK1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01021-2)
Host
Rabbit
Contents
IRAK Antibody is supplied in PBS containing 0.02% sodium azide.
Clonality
Polyclonal
Isotype
IgG
Immunogen
Anti-IRAK antibody 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.
Cross-reactivity
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.
Reactive Species
A01021-2 is reactive to IRAK1 in Human, Mouse, Rat
Observed Molecular Weight
68 kDa
Calculated molecular weight
76.5 kDa
Background of IRAK1
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).
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A01021-2 is guaranteed for ELISA, IP, IF, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
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. Optimal dilutions for each application should be determined by the researcher.
Validation Images & Assay Conditions
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Western Blot Validation in Human Cell Lines
Loading: 15 μg of lysates per lane.
Antibodies: IRAK A01021-2 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.
Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines
Loading: 15 μg of lysates per lane.
Antibodies: IRAK A01021-2 (1 μg/mL), IRAK 64-231 (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.
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Western Blot Validation with Recombinant Protein
Loading: 30 ng of human IRAK recombinant protein per lane.
Antibodies: IRAK A01021-2 (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.
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Species Activity in Mouse and Rat Cell Lines
Loading: 15 μg of lysates per lane.
Antibodies: IRAK A01021-2 (1 μg/mL,), 1h incubation at RT in 5% NFDM/TBST.
Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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Immunofluorescence Validation of IRAK in Human HeLa Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa Cells labeling IRAK with A01021-2 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
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Immunocytochemistry Validation of IRAK in Human HeLa Cells
Immunocytochemical analysis of HeLa cells using anti-IRAK antibody (A01021-2) 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.
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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 cell
extracts, untreated or treated with phosphatase as described above, were analysed for slower migrating forms of IRAK-1 by Western blotting with
anti-IRAK-1 (A01021-2). Ubiquitination was specifically detected by IRAK-1 immunoprecipitation followed by Western blotting with anti-HA antibodies.
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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.
Specific Publications For Anti-IRAK IRAK1 Antibody (A01021-2)
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