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Product Info Summary
| SKU: | A01247-1 |
|---|---|
| Size: | 0.1 mg |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, IF, ICC, WB |
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Product info
Product Name
Anti-IRAK-4 Antibody
SKU/Catalog Number
A01247-1
Size
0.1 mg
Form
Liquid
Description
Boster Bio Anti-IRAK-4 Antibody (Catalog # A01247-1). Tested in ELISA, WB, ICC, IF applications. This antibody reacts with Human.
Storage & Handling
IRAK-4 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-4 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01247-1)
Host
Rabbit
Contents
IRAK-4 Antibody is supplied in PBS containing 0.02% sodium azide.
Clonality
Polyclonal
Isotype
IgG
Immunogen
Anti-IRAK4 antibody 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.
Cross-reactivity
IRAK-4 antibody is predicted to not cross-react with other members of the IRAK protein family.
Reactive Species
A01247-1 is reactive to IRAK4 in Human
Observed Molecular Weight
68 kDa
Calculated molecular weight
51.5 kDa
Background of IRAK4
Interleukin-1 (IL-1) and lipopolysaccharide (LPS) induces cellular responses through IL-1 receptor (IL-1R) and Toll-like receptors (TLR). IL-1R-associated kinases (IRAK, IRAK2, and IRAK-M) regulate the activation of NF-κB and MAP kinase (MAPK) by IL-1R/TLR. A novel member in the IRAK/Pelle family was recently identified and designated IRAK-4. Overexpression of IRAK-4 activates NF-κB and MAPK pathways. IRAK-4 interacts with and phosphorylates IRAK-1. IRAK-4-deficient animals are completely resistant to the challenge with LPS. Animals and humans lacking IRAK-4 are impaired in their responses to viral and bacterial challenges. Members in IRAK/Pelle family play a central role in IL-1R/TLR mediated inflammatory responses and in innate immunity.
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
A01247-1 is guaranteed for ELISA, 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; 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. 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: IRAK4 A01247-1 (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: IRAK4-A01247-1 (1 μg/mL), IRAK4-24-025 (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.
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Western Blot Validation in Human HeLa Cell Lines
Loading: 15 μg of lysates per lane.
Antibodies: IRAK4 A01247-1, 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
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Immunocytochemistry Validation of IRAK4 in K562 Cells
Immunocytochemical analysis of K562 cells using anti-IRAK4 antibody (A01247-1) 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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Immunofluorescence Validation of IRAK4 in K562 Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 Cells labeling IRAK4 with A01247-1 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
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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.
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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 from ProSci, Inc.), 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.
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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.
Specific Publications For Anti-IRAK-4 Antibody (A01247-1)
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