Toll-like receptor adapter molecule 2

Other Nomenclature: Transmembrane emp24 protein transport domain containing 7; Toll like receptor adapter Molecule 2,gp24, TICAM2

 TOLL receptor signaling involved several intermediary proteins including TRAM, TRAF, MyD88, TRIF and others.  Toll-like receptor 4 (TLR4) signals the induction of transcription factor IRF3-dependent genes from the early endosome via the adaptor TRAM. A splice variant of TRAM, TAG or TRAP3b known as TRAM adaptor with GOLD domain, has golgi dynamics domain coupled to TRAM’s TOLL –interleukin 1 receptor domain.  Once cells are stimulated with lipopolysaccharide, TRAM and TIRAP localized to late endosomes positive for the GTPase Rab7a.  TIRAP3b inhibits the activation of IRF3 by LPS by displacing adapter TRIF from TRAM and induced the production of chemokine CCL5 (RANTES) but not interleukin 8 in human peripheral blood mononuclear cells (1).   TIRAP is therefore an example of a specific inhibitor of the adaptor MyD88-independent pathway activated by TLR4.

The TIRAP3b locus represents naturally occurring read-through transcription between the neighboring trans-membrane emp24 protein transport domain containing 7 (TMEd7) and toll-like receptor adaptor molecule (TICAM2) genes.  Alternate splicing results in multiple transcripts variants, one of which codes for a fusion protein that shares sequence identity with the product of each individual gene.  The fusion product negatively regulates the adapter MyD88-indepenedent toll-like receptor 4 pathway.  The variant 2 uses an alternate splice site that causes an early stop codon in the 3’-coding region, compared to variant 1. The variant 2 has a shorter C-terminus and shares no sequence identity with downstream TICAM2 gene product compared to isoform 1.  TIRAP3b sequence is also present in a trans-membrane emp24 protein transport domain contain 7 (emp24), a glycoprotein associated with cellular cargo transport complexes, COPI and COPII coated structures throughout cytoplasm.  Emp24 associate with other cellular transport proteins like gp23, gp27 in heteromeric complexes (2). TIRAP3b is a single-pass type I membrane protein and belongs to type emp24/gp25L family.  The TIRAP3b gene is located on chromosome 5q21-22 and exhibited significant single nucleotide polymorphism.

TIRAP3b-selective antibodies were generated against peptides form unique region from the TIAP3b protein with in aa 140-180.  The epitope is conserved in TIRAD, TICAM and emp24 and is common in several other species beside human but was unique to TIRAP3b locus.  The TIRAP3b antibodies are affinity purified over immobilized antigen based affinity chromatography and fully characterized for their listed applications and cross reactivity.  The TIRAP3b antibodies are also available as FITC-conjugates, other types conjugates can be ordered at a nominal charge. FabGennix Int. Inc., also provides western blot positive controls and antigenic blocking peptide for this antibody.  Please inquire for pricing and availability.  FabGennix International Inc., has produced antibodies to a wide range of targets, for a complete listing please visit www.FabGennix.com.  FabGennix antibodies, reagents and kits are now available for sale via our worldwide distribution centers. 

Immunogen:            Synthetic peptide from amino acids 147-166  corresponding to human TIRAP3b longfrom protein, the corresponding amino acids are (mes acv sih eal ksv idy qt).  The synthetic peptide was post-synthetically modified to achieve highest antigenicity before used for coupling to KLH using heterobifunctional cross linker for immunogen preparation. 

Concentration:         TIRAP-101AP and TIRAP-FITC antibody concentrations vary between 0.6-0.65 mg/ml in antibody stabilization buffer. 

Applications:        Antibody TIRAP-101AP is ideal for WB applications for detection of endogenous TIRAP3b and its cognate  gene products including short form.  The use of this antibody in other applications has not yet delineated. Investigators using this antibody in applications other than listed here can request for a complimentary sample. The dilutions for this antibody is for reference only, investigators are expected to determine the optimal conditions.    WB: > 1:250; IMM: n.d; IHC = nd.

Reactivity:                This antibody detects a band of approximately 49kDa, and 25kDa  protein in PC-TIRAP3b samples.

Protocols:            Standard protocol for various applications (WB; IMM and IHC) of this antibody is provided with the product specification sheet, however, FabGennix Inc.  strongly recommends investigators to optimize conditions for use of this antibody in their laboratories. 

 

For research use only, not for diagnostic or therapeutic use.