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IRAK4 Antibody

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Western Blot: IRAK4 Antibody [NB500-597] - Analysis of IRAK-4 in 30 ugs of NIH 3T3 cell lysate using this antibody at 1:500 dilution.
Western Blot: IRAK4 Antibody [NB500-597] - Analysis of IRAK-4 in 30 ugs of NIH 3T3 cell lysate using this antibody at 1:500 dilution.

Product Details

Summary
Reactivity Hu, Mu, RtSpecies Glossary
Applications WB, Flow, IP
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml

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IRAK4 Antibody Summary

Immunogen
This antibody was developed against a mixture of synthetic peptides containing amino acids 38-54 and 120-136 of mouse IRAK-4.
Localization
Cell Membrane
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
IRAK4
Purity
Protein G purified
Innovator's Reward
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Applications/Dilutions

Dilutions
  • Western Blot 1:1000 - 1:2000
  • Flow Cytometry
  • Immunoprecipitation
Application Notes
Immunoprecipitation: see Pacquelet et al (2007) for details.
Control
NIH 3T3 Lysate (NB800-PC8)
IRAK4 Lysate (NBL1-12028)
Publications
Read Publications using
NB500-597 in the following applications:

  • WB
    4 publications

Reactivity Notes

Rat reactivity reported in multiple pieces of scientific literature

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Purity
Protein G purified

Alternate Names for IRAK4 Antibody

  • EC 2.7.11
  • EC 2.7.11.1
  • interleukin-1 receptor-associated kinase 4
  • IPD1
  • IRAK4
  • IRAK-4
  • NY-REN-64
  • REN64
  • Renal carcinoma antigen NY-REN-64

Background

The Toll-like receptor (TLR) family in mammal comprises a family of transmembrane proteins characterized by multiple copies of leucine rich repeats in the extracellular domain and IL-1 receptor motif in the cytoplasmic domain. Like its counterparts in Drosophila, TLRs signal through adaptor molecules (1). IL-1 receptor-associated kinases (IRAKs) are important mediators in the signal transduction of Toll/IL-1 receptor (TIR) family members (2). The cytoplasmic domains of TIR proteins interact with the adapter protein, MyD88. MyD88 then recruits IRAKs (IRAK-1, -2, and M), which in turn interact with other adapter molecules, such as TRAF6 to activate NF-kB and MAPK pathways. Recently, a new member of this family, IRAK-4 has been identified (3). IRAK-4 may act as an upstream activator of IRAK-1. IRAK-4 is important for LPS activation of TLRS. Mice lacking IRAK-4 are resistant to lethal doses of LPS and are also severely impaired in their responses to viral and bacterial challenges (4, 5).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Western Blot: IRAK1 Antibody [NBP1-77068] - Analysis of IRAK in THP-1 (THP) and HeLa (HL) whole cell lysates using NBP1-77068 at 1:2000 dilution.
Immunocytochemistry/Immunofluorescence: IRAK1 Antibody [NBP1-77068] - HeLa cells.
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Western Blot: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using 2 ug/ml on (A) human intestine and 6 ug/ml on (B) mouse intestine and C) rat intestine lysate.
Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Human skin stained with at 5 ug/ml, peroxidase-conjugate and DAB chromogen. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.
Flow (Intracellular): TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using PE conjugate of NBP2-27149. An intracellular stain was performed on Jurkat cells with TLR4 antibody (76B357.1) NBP2-27149PE (blue) and an isotype control MAB004 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to phycoerythrin.
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1 µg/lane of Recombinant Human TNF-alpha was resolved by SDS-PAGE with silver staining, under reducing (R) conditions, showing a band at 17 kDa.
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Western Blot: NFkB p105/p50 Antibody (2J10D7) [NB100-56583] - Analysis of p50 in HeLa lysate in the A) absence and B) presence of immunizing peptide using p50 antibody at 5 ug/ml.
Immunohistochemistry-Paraffin: NFkB p105/p50 Antibody (2J10D7) [NB100-56583] - IHC-P of rabbit aorta using NB100-56583. Secondary antibody: anti-mouse histofine ( Nisherei Bioscience Inc. ref: 414131F). Development: DAB (Dako ref: K346811-2) and counterstained with Hematoxylin. Submitted via verified customer review
Flow Cytometry: NFkB p105/p50 Antibody (2J10D7) [NB100-56583] - An intracellular stain was performed on U937 cells with NFkB p105/p50 [2J10D7] Antibody NB100-56583C (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 650.
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TRAF6 Inhibitor Peptide Set [NBP2-26506] - The TRAF6 inhibitory peptide inhibits osteoclast ifferentiation: RAW264.7 cells were plated on 24-well plates and stimulated with RANKL and M-CSF and incubated in the presence of different concentrations of control peptide (PTD) or the TRAF6. The TRAF6 at the concentration above 30 uM suppressed osteoclast differentiation.
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Western Blot: TLR2 Antibody (TL2.1) [NB100-56722] - Using the Biotin direct conjugate. Detection of TLR2 mRNA in multiple cell lines. No detection was observed in L363 or Fravel cell lines. [PMID: 23593278]
Flow (Cell Surface): TLR2 Antibody (TL2.1) [NB100-56722] - Flow (Cell Surface): - Azide Free [NB100-56726] - Analysis using the PE conjugate of NB100-56726. Staining of TLR2 on stable transfected cell line using this antibody at 1 ug/10^6 cells. Green: isotype control. Red: anti-TLR2 antibody. Image using the Azide Free form of this antibody.
Flow Cytometry: TLR2 Antibody (TL2.1) [NB100-56722] - Analysis using Azide Free version of NB100-56722. Intracellular TLR2 in 10^6 PBMs using 2 ug of TLR2 antibody. Shaded histogram: cells without antibody. Green: isotype control. Red: TLR2 antibody. Goat anti-mouse IgG FITC secondary antibody.
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Western Blot: TLR3 Antibody (40C1285.6) [NBP2-24875] - Analysis of TLR3 in lysates from A) untransfected 293, B) 293 cells transfected with human TLR3 cDNA, C) human intestine, D) placenta, E) heart and F) ovary using TLR3 antibody at 3 ug/ml. Goat anti-mouse HRP conjugate was used as secondary.
Immunohistochemistry-Paraffin: TLR3 Antibody (40C1285.6) [NBP2-24875] - Tissue section of mouse intestine using TLR3 antibody (clone 40C1285.6) at 1:500 dilution with HRP-DAB detection and hematoxylin counterstaining. The representative image shows a punctate staining of the ER and endosomes in a subset of cells in Peyer's patches (organized lymphoid nodules) in the tested section.
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Immunocytochemistry/Immunofluorescence: TLR7 Antibody [NBP2-24906] - Analysis using the Azide Free version of NBP2-24906. Staining of Raw 246.7 cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red). Image objective 40x. An antibody dilution of 1:10 was used.
Flow (Intracellular): TLR7 Antibody [NBP2-24906] - Analysis using the FITC conjugate of NBP2-24906. Staining of TLR7 in human monocytes using this antibody at 0.5 ug/10^6 cells. Shaded histogram represents cells alone, green represents rabbit IgG isotype control, red represents anti-TLR7 antibody.
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Western Blot: TLR9 Antibody (26C593.2) [NBP2-24729] - Analysis of TLR9 in A) human PBMC, B) human intestine, C) mouse intestine, and D) rat intestine tissue lysates using Azide free TLR9 antibody at a dilution of 3 ug/mL.
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IKK gamma
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NBP2-26504
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In vivo assay: IKK gamma Inhibitor Peptide Set [NBP2-26504] - NBD peptide blocks constitutive NF-kB activation in human multiple myeloma cells. U266 cells were treated with 100 uM of control (A & B) or NBD peptide (C & D) for 12 hr, cytospun, plated on glass slides, air dried for 1 hr at room temperature and fixed with cold acetone. Slides were blocked with 5% normal goat serum for 1 hr and then incubated with rabbit polyclonal anti-human p65 antibody (A & C) followed by Ig-Alexa 594 second step. In control peptide treated cells, p65 translocates to nucleus (A), whereas NBD peptide prevents translocation of p65 into the nucleus (C). B & D: Nuclear staining with DNA binding dye.
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p38 alpha Antibody
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Western blot shows recombinant human p38 beta, p38 gamma, p38δ and Recombinant Human Active p38 alpha (Catalog # 5477-KS) (2 ng/lane). PVDF membrane was probed with 0.5 µg/mL Rabbit Anti-Human/Mouse/Rat p38 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8691) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and NIH‑3T3 mouse embryonic fibroblast cell line, loaded at 0.2 mg/mL. A specific band was detected for p38 alpha at approximately 43 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat p38 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8691). This experiment was conducted under reducing conditions and using the12-230 kDa separation system.
Orexin B, p38 alpha, and Integrin  beta 1 were detected in perfusion fixed frozen sections of mouse brainstem using Human Orexin B Monoclonal Antibody (red; Catalog # MAB734), Rabbit Anti-Human/Mouse/Rat p38 alpha Antigen Affinity-purified Polyclonal Antibody (green; Catalog # AF8691), and Mouse Integrin  beta 1 Antigen Affinity-purified Polyclonal Antibody (blue; Catalog # AF2405). Tissue was incubated with primary antibodies overnight at 4 °C. Tissue was stained using NorthernLights™ 493-conjugated Anti-Rabbit IgG Secondary Antibody (green; Catalog # NL006), NorthernLights 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007), and NorthernLights 637-conjugated Anti-Goat IgG Secondary Antibody (blue; Catalog # NL002). The image of Integrin beta 1 is pseudo-colored for presentation. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
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JNK1 Antibody
(228601) [Unconjugate
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Western blot shows lysates of CHP-100 human neuroblastoma cell line, C6 rat glioma cell line, and C2C12 mouse myoblast cell line. PVDF membrane was probed with 1 µg/mL Mouse Anti-Human/Mouse/Rat JNK1 Monoclonal Antibody (Catalog # MAB17761) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band for JNK1 was detected at approximately 46 kDa and 54 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
JNK1 was detected in immersion fixed MCF‑7 human breast cancer cell line using Mouse Anti-Human/Mouse/Rat JNK1 Monoclonal Antibody (Catalog # MAB17761) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Western blot shows recombinant human JNK1, JNK2, and JNK3 (1 ng/lane). PVDF membrane was probed with 1 µg/mL Mouse Anti-Human/Mouse/Rat JNK1 Monoclonal Antibody (Catalog # MAB17761) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band for JNK1 was detected at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
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IRAK2 Antibody
NB100-56447
Western Blot: IRAK2 Antibody [NB100-56447] - Western blot analysis of IRAK-2 in 15 ugs of Jurkat cell lysate using NB100-56447 at 1 ug/ml.
Western Blot: IRAK2 Antibody [NB100-56447] - Analysis of IRAK-2 in A) human K562 and B) mouse RAW lysate using IRAK-2 antibody at 3 ug/ml.
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TRIF/TICAM1
Antibody - Unpurified
NB120-13810
Western Blot: TRIF/TICAM1 Antibody [NB120-13810] - Analysis of TRIF in mouse spleen probed with TRIF antibody, in A: presence and B: absence of immunizing peptide, at 1:1000.
Immunocytochemistry/Immunofluorescence: TRIF/TICAM1 Antibody [NB120-13810] - TRIF antibody was tested in A431 cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red). Image objective 40X. An antibody dilution of 1:10 was used.
Western Blot: TRIF/TICAM1 Antibody [NB120-13810] - Analysis of TRIF in mouse A: Intestine. B: Spleen. C: RAW lysate using TRIF antibody at 1:1000 dilution.
(26 Publications)
Species: Hu, Mu, Rt, Po
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IRAK3 Antibody
NBP1-76782
Western Blot: IRAK3 Antibody [NBP1-76782] - Analysis of IRAK3 in mouse spleen (M) and rat liver (R) tissue lysates with IRAK3 antibody using this antibody at 1 ug/ml.
Immunohistochemistry-Paraffin: IRAK3 Antibody [NBP1-76782] - Staining of human spleen tissue with antibody at 5 ug/mL.
Immunohistochemistry-Paraffin: IRAK3 Antibody [NBP1-76782] - Immunohistochemical staining of rat liver cells using IRAK3 antibody at 2 ug/ml.
Species: Hu, Mu, Rt
Applications: WB, ELISA, IHC, IHC-P, IF
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Publications for IRAK4 Antibody (NB500-597)(11)

We have publications tested in 2 confirmed species: Mouse, Rat.

We have publications tested in 1 application: WB.


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Showing Publications 1 - 10 of 11. Show All 11 Publications.
Publications using NB500-597 Applications Species
Lee YS, Park JS, Jung SM et al. Inhibition of lethal inflammatory responses through the targeting of membrane-associated Toll-like receptor 4 signaling complexes with a Smad6-derived peptide EMBO Mol Med 2015 Mar 12 [PMID: 25766838] (WB, Mouse) WB Mouse
Valencia Pacheco GJ, Pinzon Herrera F, Cruz Lopez JJ et al. Expression and activation of intracellular receptors TLR7, TLR8 and TLR9 in peripheral blood monocytes from HIV-infected patients. Colomb Med (Cali). 2013 Jun 30 [PMID: 24892454] (WB) WB
Mirza MK, Yuan J, Gao XP et al. Caveolin-1 deficiency dampens Toll-like receptor 4 signaling through eNOS activation. Am J Pathol. 2010 May [PMID: 20304961]
Chen Yong, Liu Haizhong, Liu Zuojin et al. Blockade of inducible costimulator pathway to prevent acute rejection in rat liver transplantation. Am J Surg. 2009 Aug [PMID: 19628063] (Rat) Rat
Xu Fa Liang, You Hai Bo, Li Xu Hong et al. Glycine attenuates endotoxin-induced liver injury by downregulating TLR4 signaling in Kupffer cells. Am J Surg. 2008 Jul [PMID: 18565339] (Mouse) Mouse
Pathak SK, Basu S, Basu KK et al. Direct extracellular interaction between the early secreted antigen ESAT-6 of Mycobacterium tuberculosis and TLR2 inhibits TLR signaling in macrophages. Nat Immunol. 2007 Jun [PMID: 17486091]
Pacquelet S, Johnson JL, Ellis BA et al. Cross-talk between IRAK-4 and the NADPH oxidase. Biochem J. 2007 May 1 [PMID: 17217339]
Liu Zuo-jin, Yan Lu-nan, Li Shen-wei et al. Glycine blunts transplantative liver ischemia-reperfusion injury by downregulating interleukin 1 receptor associated kinase-4. Acta Pharmacol Sin. 2006 Nov [PMID: 17049125] (WB, Rat) WB Rat
Choi KC, Lee YS, Lim S et al. Smad6 negatively regulates interleukin 1-receptor-Toll-like receptor signaling through direct interaction with the adaptor Pellino-1. Nat Immunol. 2006 Oct [PMID: 16951688]
Hatao F, Muroi M, Hiki N et al. Prolonged Toll-like receptor stimulation leads to down-regulation of IRAK-4 protein. J Leukoc Biol. 2004 Oct [PMID: 15258191] (WB) WB
Show All 11 Publications.

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WB Video Protocol

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Control Lysate(s)

NIH 3T3 Whole Cell Lysate
IRAK4 Overexpression Lysate (Native)

Secondary Antibodies

 

Isotype Controls

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Check out the latest blog posts on IRAK4.
IRAK4: The "master IRAK" critical for initiating immune responses
IRAK4, also known as Interleukin-1 receptor-associated kinase 4, is a serine/threonine-protein kinase that plays a critical role in initiating innate and adaptive immune responses against foreign pathogens. It activates NF-kappaB in both Toll-like rec...  Read full blog post.

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Bioinformatics

Gene Symbol IRAK4
Entrez
Uniprot