TRIF/TICAM1 Antibody (567212) [Unconjugated]

Western blot shows lysates of Ramos human Burkitt's lymphoma cell line and Raji human Burkitt's lymphoma cell line. PVDF Membrane was probed with 1 µg/mL of Mouse Anti-Human TRIF/TICAM1 Monoclonal Antibody (Catalog # MAB6216) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for TRIF/TICAM1 at approximately 113 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

MyD88 is required for inflammasome sensing of HIV and HCV.Knockdowns of the TLR adaptors MyD88 (A, B) and TICAM-1 (TRIF, E, F) were generated and confirmed by previously described RNA interference techniques. (*) denotes comparisons with p≤0.05 compared to the mock transfected cells. Monocytes in which MyD88 was knocked down were cultured with HIVBaL (solid bars) or HCVSubject 180 (hatched bars) and pro-IL-1 beta mRNA transcription measured at 6 h (C, G) and IL-18 secretion measured at 24 h (D, H). Shown are the relative production of pro-IL-1 beta mRNA and IL-18 in MyD88 (C, D) or TICAM-1 (G, H) knockdown monocytes normalized to mock transfected monocytes (no siRNA) stimulated with the same viruses. Bars represent the mean ± S.D. for n = 6–9 independent transfection experiments, (**) denotes comparisons with p≤0.001 compared to the scramble siRNA transfected cells. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1004082), licensed under a CC-BY license. Not internally tested by R&D Systems.

Differential importance of endosomal TLRs in inflammasome sensing of HIV and HCV.Monocytes were cultured without stimulation or with HIVBaL or HCVSubject 180 and IL-1 beta , IFN alpha and measured at multiple timepoints. Fold change in expression of IL-1 beta (black bar) and IFN alpha (grey bar) following HIVBaL or HCVSubject 180 relative to stimulation with media alone is shown at 6 h (A). Functional knockdowns of the endosomal located TLRs in monocytes were generated by RNA interference. Monocytes were mock transfected or transfected with either siRNA targeting TLR7 or a non-targeting sequence (Scramble). After 24 h, cell lysates were prepared and efficacy of knockdown determined by (B) western blot and (C) qRT-PCR. (D) Specificity of the TLR7 siRNA was confirmed by qRT-PCR using primers for TLR3, TLR7, TLR8, and TLR9. (*) denotes comparisons with p≤0.05 compared to the mock transfected cells. Monocytes in which endosomal TLR knockdown were generated were cultured with HIVBaL (solid bars) or HCVSubject 180 (hatched bars) and pro-IL-1 beta mRNA transcription measured at 6 h (E–H) and IL-18 secretion measured at 24 h (I–L). Shown are the relative production of pro-IL-1 beta mRNA and IL-18 in TLR8 (E, I), TLR7 (F, J), TLR3 (G, K) and TLR9 (H, L) knockdown monocytes normalized to mock transfected monocytes (no siRNA) stimulated with the same viruses. Bars represent the mean ± S.D. for n = 6–9 independent transfection experiments, (**) denotes comparisons with p≤0.05 and (***) denoted p≤0.001 compared to the scramble siRNA transfected cells. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1004082), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB
• Clone: 567212
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

TRIF/TICAM1 Antibody (567212) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human TRIF/TICAM1
  Met474-Pro618
  Accession # Q8IUC6
• Specificity: Detects human TRIF/TICAM1 in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant human (rh) TRIF (aa 29‑204), rhTRAM, rhTRIM, rhTRIM5, rhTRIM21, or rhTRIM32 is observed.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: TICAM1
• Purity Statement: IgM-specific Affinity-purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for TRIF/TICAM1 Antibody (567212) [Unconjugated]

• 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

TRIF (TIR domain-containing adaptor inducing IFN-beta ; also TICAM1) is a 105-110 kDa cytoplasmic adaptor molecule that mediates Toll receptor signaling. It is widely expressed and associates with both TLR3 and TLR4. Relative to TLR3, TRIF appears to activate IRF-3, -4, and -7, as well as NFkB and FADD. Its action on FADD is through RIP1, and this induces apoptosis. Human TRIF is 712 amino acids (aa) in length. It contains three TRAF6 bonding motifs (aa 84-91, 248-255 and 299-309), one TIR domain (aa 390-460), a Pro-rich region (aa 614-678), and an overlapping RHIM domain (aa 661-699). The molecule is reported to form a homodimer. There are multiple potential isoform variants. One shows a 23 aa substitution for aa 31-162 accompanied by a Pro substitution for aa 633-660, a second shows a 44 aa substitution for aa 352-712, and a third shows a 38 aa substitution for aa 271-712. Over aa 474-618, human TRIF shares 53% aa identity with mouse TRIF.

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 (MAB6216)(2)

Publications using MAB6216

• M Qadri, GD Jay, LX Zhang, W Wong, AM Reginato, C Sun, TA Schmidt, KA Elsaid Recombinant human proteoglycan-4 reduces phagocytosis of urate crystals and downstream nuclear factor kappa B and inflammasome activation and production of cytokines and chemokines in human and murine macrophages Arthritis Res. Ther., 2018-08-29;20(1):192. 2018-08-29 [PMID: 30157934] (Western Blot, Human)
• Chattergoon M, Latanich R, Quinn J, Winter M, Buckheit R, Blankson J, Pardoll D, Cox A HIV and HCV activate the inflammasome in monocytes and macrophages via endosomal Toll-like receptors without induction of type 1 interferon. PLoS Pathog, 2014-05-01;10(5):e1004082. 2014-05-01 [PMID: 24788318] (Western Blot, Human)

Bioinformatics

• Gene Symbol: TICAM1
• Uniprot:
  • Q8IUC6(Human)