IRAK1 Polyclonal antibody

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  WB analysis of HEK-293 using 10478-2-AP

  WB result of IRAK1 antibody (10478-2-AP; 1:2000; incubated at room temperature for 1.5 hours) with sh-Control and sh-IRAK1 transfected HEK-293 cells.

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  WB analysis using 10478-2-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10478-2-AP (IRAK1 antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.

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  WB analysis using 10478-2-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10478-2-AP (IRAK1 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis of PC-12 using 10478-2-AP

  PC-12 cells were subjected to SDS PAGE followed by western blot with 10478-2-AP (IRAK1 antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

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  WB analysis of RAW 264.7 using 10478-2-AP

  RAW 264.7 cells were subjected to SDS PAGE followed by western blot with 10478-2-AP (IRAK1 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.

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  WB analysis of PC-13 using 10478-2-AP

  PC-13 cells were subjected to SDS PAGE followed by western blot with 10478-2-AP (IRAK1 Antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 10478-2-AP

  IP result of anti-IRAK1 (IP:10478-2-AP, 4ug; Detection:10478-2-AP 1:600) with HeLa cells lysate 2800ug.

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  IHC staining of human lung cancer using 10478-2-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 10478-2-AP (IRAK1 antibody) at dilution of 1:600 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 10478-2-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 10478-2-AP (IRAK1 antibody) at dilution of 1:600 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human placenta using 10478-2-AP

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 10478-2-AP (IRAK1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse ovary using 10478-2-AP

  Immunohistochemical analysis of paraffin-embedded mouse ovary tissue slide using 10478-2-AP (IRAK1 antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of rat intestine using 10478-2-AP

  Immunohistochemical analysis of paraffin-embedded rat intestine tissue slide using 10478-2-AP (IRAK1 antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of rat ovary using 10478-2-AP

  Immunohistochemical analysis of paraffin-embedded rat ovary tissue slide using 10478-2-AP (IRAK1 antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 10478-2-AP

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using IRAK1 antibody (10478-2-AP) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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  FC experiment of HeLa using 10478-2-AP

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human IRAK1 (10478-2-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

• Featured Product
• KD/KO Validated

IRAK1 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Cat No. 10478-2-AP

Publications(38)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (1)

Applications

WB, IHC, IF/ICC, FC (Intra), IP, RIP, ELISA

Synonyms

pelle, IRAK-1, IRAK 1, IRAK, Interleukin-1 receptor-associated kinase 1

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Tested Applications

• Positive WB detected in: A549 cells, PC-13 cells, PC-12 cells, RAW 264.7 cells, HEK-293 cells, HeLa cells, Jurkat cells, K-562 cells, MCF-7 cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human lung cancer tissue, human placenta tissue, mouse ovary tissue, rat intestine, rat ovary tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:3000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:300-1:1200
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 2 publications below
• WB: See 37 publications below
• IHC: See 7 publications below
• IF: See 2 publications below
• RIP: See 1 publications below

Product Information

10478-2-AP targets IRAK1 in WB, IHC, IF/ICC, FC (Intra), IP, RIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, sheep
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: IRAK1 fusion protein Ag0728
• Full Name: interleukin-1 receptor-associated kinase 1
• Calculated Molecular Weight: 77 kDa
• Observed Molecular Weight: 68-80 kDa
• GenBank Accession Number: BC014963
• Gene Symbol: IRAK1
• Gene ID (NCBI): 3654
• RRID: AB_2126032
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: P51617
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Interleukin-1 receptor-associated kinases (IRAKs) are a unique family of death domain containing protein kinases that play a key role in initiating innate immune response against foreign pathogens. They are involved in Toll-like receptor (TLR) and interleukin-1 receptor (IL-1R) signaling pathways. IRAK1 is the first member of this kinase family. Upon ligand binding to TLR/IL-1R, IRAK1 is recruited by MYD88 to the receptor-signaling complex, the association leads to IRAK1 phosphorylation by IRAK4 and subsequent autophosphorylation and kinase activation. Hyper-phosphorylated IRAK1 then disengages from the receptor complex, and forms a cytosolic IRAK1-TRAF6 complex. TRAF6 then interacts with TAK and TAB, resulting in eventual activation of the NF-κB and MAPK pathways. Phosphorylated IRAK1 also undergoes ubiquitin-mediated degradation or sumoylation, which results in nuclear translocation and transcriptional activation of inflammatory target genes. (PMID: 17890055; 12620219)

Publications

Species	Application	Title
human	WB	Blood; Targeting of microRNA-142-3p in dendritic cells regulates endotoxin-induced mortality.; Authors - Sun Yaping Y
mouse	WB	Cell Rep; METTL3-mediated m6A mRNA methylation regulates neutrophil activation through targeting TLR4 signaling; Authors - Shuhua Luo
mouse	WB,IHC	J Crohns Colitis; A Molecular Targeted Immunotherapeutic Strategy for Ulcerative Colitis via Dual-targeting Nanoparticles Delivering miR-146b to Intestinal Macrophages.; Authors - Feihong Deng
mouse	WB	Cell Mol Immunol; The deubiquitinase OTUB1 augments NF-κB-dependent immune responses in dendritic cells in infection and inflammation by stabilizing UBC13.; Authors - Floriana Mulas
human	WB	Cell Mol Immunol; Virus-inducible IGFALS facilitates innate immune responses by mediating IRAK1 and TRAF6 activation.; Authors - Gang Xu
mouse	WB	Mol Cell Proteomics; Quantification of Dynamic Protein Interactions and Phosphorylation in LPS Signaling Pathway by SWATH-MS; Authors - Xiurong Wu