TRIF/TICAM1 Antibody

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.

Simple Western: TRIF/TICAM1 Antibody [NB120-13810] - Image shows a specific band for TRIF in 0.5 mg/mL of Raji lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu, Mu, Rt, Po
• Applications: WB, Simple Western, IB, ICC/IF, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

TRIF/TICAM1 Antibody Summary

• Immunogen: Synthetic peptide made to an internal portion of the mouse TRIF protein (between amino acids 100-180) [UniProt Q80UF7].
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: TICAM1
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:500
  • Simple Western 1:100
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:10-1:100
  • Immunoprecipitation
• Application Notes: In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
  The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. Use in Immunoprecipitation reported in scientific literature (PMID 28007523). Use in Immunoblotting reported in scientific literature (PMID 27043414)..
• Theoretical MW: 76 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  Mouse Spleen Whole Tissue Lysate (Adult Whole Normal)
  TRIF/TICAM1 Overexpression Lysate

Reactivity Notes

Human reactivity reported in the scientific literature (PMID: 23777661). Rat reactivity reported in scientific literature (PMID: 23447644). Porcine reactivity reported in scientific literature (PMID: 27046485).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Whole antisera
• Preservative: 0.02% Sodium Azide
• Purity: Unpurified

Alternate Names for TRIF/TICAM1 Antibody

• MGC35334
• Proline-rich, vinculin and TIR domain-containing protein B
• PRVTIRBTIR domain-containing adapter protein inducing IFN-beta
• Putative NF-kappa-B-activating protein 502H
• TICAM1
• TICAM-1TIR domain containing adaptor inducing interferon-beta
• TIR domain-containing adapter molecule 1
• TIR domain-containing adaptor molecule 1
• toll-like receptor adaptor molecule 1
• TRIF
• TRIFToll-interleukin-1 receptor domain-containing adapter protein inducinginterferon beta

Background

TICAM (Toll-interleukin 1 receptor domain (TIR)-containing adaptor molecule-1) or TRIF (Toll/IL-1 receptor domain-containing adapter inducing IFN-beta) plays an essential role in the MyD88-independent signaling of TLR3.

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.

Publications for TRIF/TICAM1 Antibody (NB120-13810)(28)

Publications using NB120-13810

• Feng Z, Yang R, Wu L et al. Atractylodes macrocephala polysaccharides regulate the innate immunity of colorectal cancer cells by modulating the TLR4 signaling pathway Onco Targets Ther [PMID: 31564895] (Mouse)
• Zhou S, Qi Q, Wang X, et al. SjHSP60 induces CD4+ CD25+ Foxp3+ Tregs via TLR4-Mal-drived production of TGF-beta in macrophages. Immunol Cell Biol. 2018 May 21 [PMID: 29697865] (WB, Mouse)
• Gao T, Zhang SP, Wang JF et al. TLR3 contributes to persistent autophagy and heart failure in mice after myocardial infarction J. Cell. Mol. Med. 2017 Sep 25 [PMID: 28945004] (WB, Mouse)
• Popiolek-Barczyk K, Piotrowska A, Makuch W, Mika J. Biphalin, a Dimeric Enkephalin, Alleviates LPS-Induced Activation in Rat Primary Microglial Cultures in Opioid Receptor-Dependent and Receptor-Independent Manners.. Neural Plast. Jun 2 2017 [PMID: 28573049] (WB, Rat)
• Kim E, Beon J, Lee S et al. Inositol polyphosphate multikinase promotes Toll-like receptor-induced inflammation by stabilizing TRAF6. Sci Adv. Apr 1 2017 [PMID: 28439546] (IP, Mouse)
• Lan X, Han X, Li Q et al. Pinocembrin protects hemorrhagic brain primarily by inhibiting toll-like receptor 4 and reducing M1 phenotype microglia Brain Behav. Immun. Dec 20 2016 [PMID: 28007523] (IP, Mouse)
• Zhang L, Xiang W, Wang G et al. IFN-beta Production During the dsRNA Response in Hepatocytes Involves Coordinated and Feed Forward Signaling Through TLR3, PkR, iNOS and Src. J. Biol. Chem. May 17 2016 [PMID: 27226571] (WB, Mouse)
• Hu Y, Cong X, Chen L et al. Synergy of TLR3 and 7 ligands significantly enhances function of DCs to present inactivated PRRSV antigen through TRIF/MyD88-NF-kB signaling pathway. Sci Rep. 2016 Apr 06 [PMID: 27046485] (WB, Porcine)
• Wang F, Alain T, Szretter KJ et al. S6K-STING interaction regulates cytosolic DNA-mediated activation of the transcription factor IRF3. Nat. Immunol. May 1 2016 [PMID: 27043414] (IB, IP, Mouse)
• Ma B, Yu J, Xie C et al. Toll-Like Receptors Promote Mitochondrial Translocation of Nuclear Transcription Factor Nuclear Factor of Activated T-Cells in Prolonged Microglial Activation. J. Neurosci. 2015 Jul 29 [PMID: 26224862] (ICC/IF, Mouse)
  
  Details:
  TRIF/TICAM1 antibody (Imgenex IMG-661) was used for ICC-IF analysis of Trif siRNA transfected primary microglial cell cultures that were isolated from Nfat1 knock-out mice pups. The assay involved 4% PFA fixation, permeabilization with Triton X-100, blocking with 10% donkey serum, use of primary antibody at 1:500 dilution and detection using Alexa labelled secondary antibody (Figure 7A).
• Vogt A, Scull MA, Friling T et al. Recapitulation of the hepatitis C virus life-cycle in engineered murine cell lines. Virology 2013 Jun 15 [PMID: 23777661] (WB, Mouse, Human)
• Haque A, Koide N, Odkhuu E et al. Mouse pyrin and HIN domain family member 1 (pyhin1) protein positively regulates LPS-induced IFN-B and NO production in macrophages. Innate Immun. 2014 Jan 1 [PMID: 23606517] (WB, Mouse)
• Tian J, Guo X, Liu XM et al. Extracellular HSP60 induces inflammation through activating and up-regulating TLRs in cardiomyocytes. Cardiovasc Res. 2013 Jun 1 [PMID: 23447644] (WB, Rat)
• Abe T, Hirasaka K, Kagawa S et al. Cbl-b Is a Critical Regulator of Macrophage Activation Associated with Obesity-Induced Insulin Resistance in Mice. Diabetes 2013 Jan 24 [PMID: 23349502]
• De Luca A, Iannitti RG, Bozza S et al. CD4(+) T cell vaccination overcomes defective cross-presentation of fungal antigens in a mouse model of chronic granulomatous disease. J Clin Invest. 2012 May 1 [PMID: 22523066] (WB)
  
  Details:
  WB (Fig 7F): EGFP-LC3-transfected RAW 264.7 cells. Cell lysates were IP'd with an antibody to Beclin-1 and then western blotted with the IMG-661 TRIF pAb. NOTE: The TRIF IMG-661 antibody recognized TRIF by WB that had been IP'd with a Beclin-1 Ab in an IP
• Seleme MC, Lei W, Burg AR et al. Dysregulated TLR3-dependent signaling and innate immune activation in superoxide-deficient macrophages from nonobese diabetic mice. Free Radic Biol Med. 2012 May 1 [PMID: 22361747] (WB)
  
  Details:
  WB (bone marrow derived mouse macrophages): Fig 6C,D.
• Zhang Z, Kim T, Bao M et al. DDX1, DDX21, and DHX36 Helicases Form a Complex with the Adaptor Molecule TRIF to Sense dsRNA in Dendritic Cells. Immunity;34(6):866-78. 2011 Jun 24. [PMID: 21703541]
• Kong X, Thimmulappa R, Kombairaju P, Biswal S. NADPH oxidase-dependent reactive oxygen species mediate amplified TLR4 signaling and sepsis-induced mortality in Nrf2-deficient mice. J Immunol. 2010 Jul 1 [PMID: 20511556] (WB, Mouse)
  
  Details:
  WB (mouse macrophages treated with LPS), Fig 4. Note: Trif was found to co-IP with TLR4.
• Shi CS, Kehrl JH. TRAF6 and A20 regulate lysine 63-linked ubiquitination of Beclin-1 to control TLR4-induced autophagy. Sci Signal. 2010 May 25 [PMID: 20501938] (WB)
  
  Details:
  WB (RAW 267.4 cells): Fig 1B (bottom panel).
• Wong SW, Kwon MJ, Choi AM et al. Fatty acids modulate Toll-like receptor 4 activation through regulation of receptor dimerization and recruitment into lipid rafts in a reactive oxygen species-dependent manner. J Biol Chem. 2009 Oct 2 [PMID: 19648648] (WB)
  
  Details:
  WB (RAW 264.7 cells treated with LPS, lauric acid or docosahexaenoic acid), Figs 1,2A-B. The expression of Trif in lipid and non-lipid raft fractions was analyzed.
• Dasu MR, Park S, Devaraj S, Jialal I. Pioglitazone inhibits Toll-like receptor expression and activity in human monocytes and db/db mice. Endocrinology. 2009 Aug [PMID: 19389833] (WB)
  
  Details:
  WB (mouse peritoneal macrophages from db/db mice): Fig 3A. Pioglitazone treatment of mice greatly reduced TRIF expression. Note: db/db mouse is a model of obesity, diabetes and dylipidemisa. They are homozygous for a point mutation in the leptin receptor
• Wang XM, Kim HP, Nakahira K et al. The heme oxygenase-1/carbon monoxide pathway suppresses TLR4 signaling by regulating the interaction of TLR4 with caveolin-1. J Immunol. 2009 Mar 15 [PMID: 19265160] (WB, IP)
  
  Details:
  Raw 264.7 cells expressing cav-l, WB: Fig 1E (left), IP: Fig 1E (right). Note: Trif was found to co-IP with TLR4 (Fig 1E left). Likewise, the Trif antibody co-IP'd TLR4 (Fig 1E right).
• Shi CS, Kehrl JH. MyD88 and Trif target Beclin 1 to trigger autophagy in macrophages. J Biol Chem. 2008 Nov 28 [PMID: 18772134] (WB)
  
  Details:
  WB (mouse RAW264.7 cells), Fig. 2F; 3B,C; 4A,B,F. MyD88 and Trif target Beclin1 to trigger autophagy in macrophages. Shi C, J Kehrl. JBCdoi:10.1074/jbc.M804478200 (2008). WB (mouse RAW264.7 cells), Fig. 2F; 3B,C; 4A,B,F.
• Krikun G, Lockwood CJ, Abrahams VM et al. Expression of Toll-like receptors in the human decidua. Histol Histopathol. 2007 Aug [PMID: 17503341] (WB, Human)
  
  Details:
  WB (first trimester human decidual cell cultures), Fig 5. Trif was detected at ~66 kDa.
• Palladino MA, Johnson TA, Gupta R et al. Members of the Toll-like receptor family of innate immunity pattern-recognition receptors are abundant in the male rat reproductive tract. Biol Reprod. 2007 Jun [PMID: 17314314] (WB, Rat)
  
  Details:
  Anitbodies cited: The following antibodies were used for WB in Fig 2 in rat testis, epididymis, and vas deferens: TLR1 (IMG-5012), TLR2 (IMG-545), TLR5 (IMG-580), TLR6 (IMG-527), TLR7 (IMG-581), TLR9 (IMG-305A), and TLR12 (IMG-5034). MYD88 (IMG-178) and T
• Nakahira K, Kim HP, Geng XH et al. Carbon monoxide differentially inhibits TLR signaling pathways by regulating ROS-induced trafficking of TLRs to lipid rafts. J Exp Med. 2006 Oct 2 [PMID: 17000866] (WB)
  
  Details:
  WB [Raw 264.7 cells treated with LPS or poly (I:C) in the absence or presence of carbon monoxide], Figs 6, 7B&C, 8C. Note: Trif was found to co-IP with TLR4 and TLR3.
• Chassin C, Goujon JM, Darche S et al. Renal collecting duct epithelial cells react to pyelonephritis-associated Escherichia coli by activating distinct TLR4-dependent and -independent inflammatory pathways. J Immunol. 2006 Oct 1 [PMID: 16982918] (WB, Mouse)
  
  Details:
  WB (mouse kidney tissue lysate), Fig 7A.
• Naiki Y, Michelsen KS, Zhang W et al. Transforming growth factor-beta differentially inhibits MyD88-dependent, but not TRAM- and TRIF-dependent, lipopolysaccharide-induced TLR4 signaling. J Biol Chem. 2005 Feb 18 [PMID: 15623538] (WB, Human)
  
  Details:
  IP/WB (Fig 3E): human dermal mircrovessel endothelial cells.

FAQs for TRIF/TICAM1 Antibody (NB120-13810). (Showing 1 - 1 of 1 FAQs).

1. Regarding the WB testing for the TRIF/TICAM1 Antibody Cat# NB120-13810, can you confirm if the RAW lysate was treated in any way? Some of the antibody references on the product page indicate samples treated with LPS. Is there also any explanation for the molecular weight observed? The signal on mouse is right above the 66 kDa maker while the testing on Raji displays a band closer to 100 kDa.
   • The RAW lysate has not been treated in the western shown here. The difference in sizes can be explained as the mouse sample that is running right at 66 KD is a regular Western Blot, however, the band that is running at 100 KD is on the Simple Western system and tends to run larger than on a regular western blot. Protein Simple is the new line of proteins analysis technology:

Bioinformatics

• Gene Symbol: TICAM1
• Entrez:
  • 106759(Mouse)
• Uniprot:
  • Q80UF7(Mouse)