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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

Order Details

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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 Whole Cell Lysate
IRAK4 Overexpression Lysate
Publications
Read Publications using
NB500-597 in the following applications:

  • WB
    5 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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Immunocytochemistry/Immunofluorescence: IRAK1 Antibody [NBP1-77068] - Immunofluorescence Validation of IRAK in Human HeLa Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa Cells labeling IRAK with NBP1-77068 at 20 ug g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
Western Blot: IRAK1 Antibody [NBP1-77068] - Western Blot Validation in Human Cell Lines. Loading: 15 ug of lysates per lane.Antibodies: IRAK NBP1-77068 (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
(1 Publication)
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(76B357.1)
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Dual RNAscope ISH-IHC: TLR4 Antibody (76B357.1) [NB100-56566] - FFPE tissue sections of human tonsil were probed for TLR4 mRNA (ACD RNAScope Probe, ACD catalog # 311281; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Mouse Monoclonal (Novus Biologicals catalog # NB100-56566) at 5ug/mL with 1 hour incubation at room temperature followed by incubation with anti-mouse IgG VisUCyte HRP Polymer Antibody (Catalog # VC001) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes.
Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using the Alexa Fluor (R) 647 conjugate of NBP2-27149. TLR4 expression on monocytes from human peripheral blood. PBMC were stained in a 2 color flow test, with CD14 PE version of this antibody and 1 ug of either isotype control (left) or TLR4-Alexa Fluor 647 (right). PPI negative, CD14+ cells were gated for analysis.
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201-LB
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NFkB p105/p50
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NB100-56583
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.
(1 Review)
(4 Publications)
Species: Hu, Rt, Rb
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NBP2-26506
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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Flow Cytometry: TLR2 Antibody (TL2.1) [NB100-56722] - Analysis using the FITC conjugate of NB100-56726. Surface staining of stable HEK293/hTLR2 cells (IML-202, red) and vector control cells (IML-200, green) using TLR2 antibody at 1 ug/10^6 cells.
Western Blot: TLR2 Antibody (TL2.1) [NB100-56722] - Expression of TLR2 was determined by western blotting. The immunoreactivity of the anti-TLR2 was confirmed with human intestinal lysate. Data are representative for analysis of >=2 independent experiments. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0060671) licensed under a CC-BY licence.
Western Blot: TLR2 Antibody (TL2.1) [NB100-56722] - Total protein from human THP-1 and U937 cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-TLR2 (NB100-56722) in 5% BSA-TBST and detected with an anti-mouse HRP secondary antibody using NovaLume chemiluminescence detection reagent (NPB2-61915).
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IL-6 [HRP]
D6050
(25 Reviews)
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(40C1285.6)
NBP2-24875
Flow Cytometry: TLR3 Antibody (40C1285.6) [NBP2-24875] - Expression of TLR3 protein on epithelial cells. HNEC (A), Detroit-562 (B) and FaDu (C) were stained intracellularly with FITC-Abs against TLR3 (open histograms) or appropriate isotype control (shaded histograms) and analyzed by flow cytometry. Representative pictures from one out of three independent experiments are shown. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0098239), licensed under a CC-BY licence.
Western Blot: TLR3 Antibody (40C1285.6) [NBP2-24875] - All the cell lines displayed a strong expression of TLR3 protein. Cell lysates were electrophoresed and blotted to PVDF membrane, which was probed with TLR3-specific antibody. To confirm the immunoreactivity of the antibody, different positive controls were included. Beta-actin was served as loading control and was used to normalize expression levels between cells. Data are representative for analysis of >=2 independent experiments. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0060671) licensed under a CC-BY licence.
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NBP2-24906
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Flow Cytometry: TLR7 Antibody [NBP2-24906] - An intracellular stain was performed on THP-1 cells with NBP2-24906 (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 5.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550.
Immunocytochemistry/Immunofluorescence: TLR7 Antibody [NBP2-24906] - Immunolocalization of TLR7 (A) and glial fibrillary acidic protein (GFAP) (B) in murine myenteric plexus. Most of the GFAP positive cells were also positive for TLR7 in the merged image (C, yellow staining). (Courtesy of Barajon I, et al., Journal of Histochemistry and Cytochemistry, 57(11): 1013-;1023, 2009.)
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TLR9 Antibody
(26C593.2)
NBP2-24729
Simple Western: TLR9 Antibody (26C593.2) [NBP2-24729] - Lane view shows a specific band for TLR9 in 0.5 mg/ml of Ramos lysate. This experiment was performed under reducing conditions using the 66-440 kDa separation system. Image using the Azide Free format of this antibody.
Flow Cytometry: TLR9 Antibody (26C593.2) [NBP2-24729] - Expression of TLR9 protein on epithelial cells. HNEC, Detroit-562 and FaDu were stained intracellularly with PE-Ab against TLR9 (open histograms) or appropriate isotype control (shaded histograms) and analyzed by flow cytometry. Representative pictures from one out of three independent experiments are shown. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0098239), licensed under a CC-BY licence.
Knockdown Validated: TLR9 Antibody (26C593.2) [NBP2-24729] - Expression of TLR-9 protein in HB cells before and after 48 hr of transfection. Beta-actin was used as an internal control. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0092748) licensed under a CC-BY licence.
(4 Reviews)
(142 Publications)
Species: Hu, Mu, Rt, Ca, Eq, Pm, Pm
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ERK2 Antibody
(191801) [Unconjugate
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Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, MCF‑7 human breast cancer cell line, U937 human histiocytic lymphoma cell line,PC‑12 rat adrenal pheochromocytoma cell line, and NIH‑3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat ERK2 Monoclonal Antibody (Catalog # MAB1230) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for ERK2 at approximately 44 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
ERK2 was detected in immersion fixed 3T3‑L1 mouse embryonic fibroblast adipose-like cell line using Mouse Anti-Human/Mouse/Rat ERK2 Monoclonal Antibody (Catalog # MAB1230) at 3 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 cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and MCF‑7 human breast cancer cell line, loaded at 0.5 mg/mL. A specific band was detected for ERK2 at approximately 43 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human/Mouse/Rat ERK2 Monoclonal Antibody (Catalog # MAB1230). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Species: Hu, Mu, Rt
Applications: WB, Simple Western, IHC, ICC, KO
IKK gamma
Inhibitor Peptide Set
NBP2-26504
In vivo assay: IKK gamma Inhibitor Peptide Set [NBP2-26504] - NBD peptide blocks constitutive NF-kB as shown by EMSA. U266 cells were treated with 100 uM of control or NBD peptide for different time periods. Nuclear extracts were isolated and checked for NF-kB-DNA binding activity.
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.
IKK gamma Inhibitor Peptide Set [NBP2-26504] - The IKK complex consists of IKKalpha + IKKbeta+ NEMO. NEMO (IKKgamma/NF-kB Essential MOdulator) is a scaffold protein required for the kinase activity of IKKalpha + IKKbeta. NBD (NEMO Binding Domain) inhibits formation of the IKK complex (IKKalpha + IKKbeta+ NEMO) by binding to NEMO and blocking IKK complex formation. In this regard, NBD is considered to be an 'IKKalpha and IKKBeta decoy' Blocking IKK complex formation suppresses downstream events in the NF-kB signaling pathway that rely on the IKK complex including phosphorylation of IkBalpha, and subsequence IkBalpha ubiquination, degradation and release of NF-kB. Hence, NF-kB is retained in the cytoplasm in its inactive state.
(23 Publications)
Species: Hu, Mu, Rt
p38 alpha [p
Thr180, p Tyr182] Ant
NB500-138
Western Blot: p38 [p Thr180, p Tyr182] Antibody [NB500-138] - Hela cell lysates that had been treated with UV or untreated (Control) showing specific immunolabeling of the -39 kDa p38 MAPK protein phosphorylated at Thr180/Tyr182.
Immunohistochemistry-Paraffin: p38 [p Thr180, p Tyr182] Antibody [NB500-138] - Immunostaining of human breast cancer tissue showing p38 when phosphorylated at Thr180/Tyr182 in red.
Western Blot: p38 [p Thr180, p Tyr182] Antibody [NB500-138] - Western blot of HeLa cell lysates that had been treated with UV or untreated (Control) showing specific immunolabeling of the ~39k p38 MAPK protein phosphorylated at Thr180/Tyr182.
(12 Publications)
Species: Hu, Mu, Rt, Ze
Applications: WB, IHC, IHC-P, Flow-IC
JNK1 Antibody
(228601) [Unconjugate
MAB17761
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.
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.
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.
(2 Reviews)
(3 Publications)
Species: Hu, Mu, Rt
Applications: WB, ICC
IRAK2 Antibody
(JG36-23)
NBP2-76843
Western Blot: IRAK2 Antibody (JG36-23) [NBP2-76843] - Western blot analysis of IRAK2 on different cell lysates using anti-IRAK2 at 1/500 dilution.Positive control:Line 1: PC-3M Line 2: A549Line 3: HepG2
Immunocytochemistry/Immunofluorescence: IRAK2 Antibody (JG36-23) [NBP2-76843] - ICC staining IRAK2 in PC-3M cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Immunohistochemistry: IRAK2 Antibody (JG36-23) [NBP2-76843] - Immunohistochemical analysis of paraffin-embedded human heart tissue using anti-IRAK2 antibody. Counter stained with hematoxylin.
Species: Hu
Applications: WB, Flow, ICC/IF, IHC, IHC-P, IP
TRIF/TICAM1
Antibody
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.
(31 Publications)
Species: Hu, Mu, Rt, Po
Applications: WB, Simple Western, IB, ICC/IF, IP
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
IL-18/IL-1F4 [HRP]
7620
IL-18/IL-1F4 [HRP]
(39 Publications)
Species: Hu
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Publications for IRAK4 Antibody (NB500-597)(12)

We have publications tested in 3 confirmed species: Human, Mouse, Rat.

We have publications tested in 1 application: WB.


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Showing Publications 1 - 10 of 12. Show All 12 Publications.
Publications using NB500-597 Applications Species
Li, Wang H, Li Z et al. The inhibition of RANKL expression in fibroblasts attenuate CoCr particles induced aseptic prosthesis loosening via the MyD88-independent TLR signaling pathway. Biochem. Biophys. Res. Commun. Jun 25 2018 [PMID: 29940143] (WB, Human) WB Human
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]
Show All 12 Publications.

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Control Lysate(s)

NIH 3T3 Whole Cell Lysate
IRAK4 Overexpression Lysate

Secondary Antibodies

 

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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