TRIF/TICAM1 Antibody - BSA Free

Western Blot: TRIF/TICAM1 Antibody [NB120-13810] - Total protein from Raw264.7 cells and mouse spleen 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-TICAM1 (NB120-13810) in blocking buffer and detected with an anti-rabbit HRP secondary antibody using NovaLume chemiluminescence detection reagent (NBP2-61915).

Immunohistochemistry-Paraffin: TRIF/TICAM1 Antibody [NB120-13810] - Analysis of a FFPE tissue section of mouse spleen using 1:200 dilution of TRIF/TICAM1 antibody. The staining was developed using HRP labeled anti-rabbit secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.

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.

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.

Knockdown Validated: TRIF/TICAM1 Antibody [NB120-13810] - siRNA was transfected at 50 nM for 48 hrs, and poly(I:C) was added at 100 ug/ml for 4 hrs before cell harvest. Negative control (NC) siRNA served as control. RNAiMAX transfection reagent was used in all the siRNA experiments. The panel shows the knockdown effect of Trif siRNA. Image collected and cropped by CiteAb from the following publication (//onlinelibrary.wiley.com/doi/full/10.1111/jcmm.13328) licensed under a CC-BY license.

TRIF/TICAM1 Antibody [NB120-13810] - Lysates of heart tissue were immunoprecipitated with anti-TLR3 antibodies (IP: TLR3), followed by SDS-PAGE and immunoblotting (IB) with indicated antibodies. IP with isotype IgG (IP: IgG) was performed as a control to exclude the non-specific binding of antibodies to cellular proteins. Green arrows indicate non-specific bands. The association between TLR3 and Trif, but not MyD88, was detectable in sham myocardium and was increased in infarct. Image collected and cropped by CiteAb from the following publication (//onlinelibrary.wiley.com/doi/full/10.1111/jcmm.13328) licensed under a CC-BY license. Anti-TLR3 antibody by WB (NB100-56571)

• Reactivity: Mu, Rt, Po
• Applications: WB, Simple Western, ICC/IF, IHC, IP, WB, KD
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

TRIF/TICAM1 Antibody - BSA Free 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: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Immunoblotting reported in scientific literature (PMID 27043414)
  • Immunocytochemistry/Immunofluorescence 1:10-1:100
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Paraffin 1:200
  • Immunoprecipitation reported in scientific literature (PMID 28007523)
  • Knockdown Validated
  • Simple Western 1:100
  • Western Blot 1 - 2 ug/ml
• 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.
• 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

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: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for TRIF/TICAM1 Antibody - BSA Free

• 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)(29)

Publications using NB120-13810

• Youn SE, Jiang F, Won HY et al. PAUF Induces Migration of Human Pancreatic Cancer Cells Exclusively via the TLR4/MyD88/NF-kappa B Signaling Pathway International journal of molecular sciences 2022-09-27 [PMID: 36232715] (IP, Human)
• Karmakar J, Mandal C Interplay Between Sialic Acids, Siglec-E, and Neu1 Regulates MyD88- and TRIF-Dependent Pathways for TLR4-Activation During Leishmania donovani Infection Frontiers in immunology 2021-03-03 [PMID: 33763070] (WB, Mouse)
• Chen YL, Shirakawa H, Lu NS et al. Impacts of fish oil on the gut microbiota of rats with alcoholic liver damage J. Nutr. Biochem. 2020-09-10 [PMID: 32920090] (WB, Rat)
• Shi D, Chen M, Liu L et al. Anti-influenza A virus mechanism of three representative compounds from Flos Trollii via TLRs signaling pathways J Ethnopharmacol 2020-01-28 [PMID: 32004628] (WB, Mouse)
• 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. 2018-05-21 [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. 2017-09-25 [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. 2017-06-02 [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. 2017-04-01 [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. 2016-12-20 [PMID: 28007523] (IP, Mouse)
• Zhang L, Xiang W, Wang G et al. #VALUE! J Biol Chem. [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. 2016-04-06 [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. 2016-05-01 [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. 2015-07-29 [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).
• 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. 2014-01-01 [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. 2013-06-01 [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 2013-01-24 [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. 2012-05-01 [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. 2012-05-01 [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. 2011-06-24 [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. 2010-07-01 [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. 2010-05-25 [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. 2009-10-02 [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. 2009-08-01 [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. 2009-03-15 [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. 2008-11-28 [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.
• Palladino M A, Johnson T A, 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. 2007-06-01 [PMID: 17314314] (WB, Rat)
• 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. 2006-10-02 [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. 2006-10-01 [PMID: 16982918] (WB, Mouse)
  
  Details:
  WB (mouse kidney tissue lysate), Fig 7A.

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)