Flow cytometric analysis of Raji cells, using TRAF1 (45D3) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
Western blot analysis of extracts from Jurkat, 293, HeLa, C2C12, and NIH/3T3 cells, using TRAF2 (C192) Antibody.
Western blot analysis of extracts from COS-7 cells, untransfected (-) or transfected with human TRAF3 construct (+) using TRAF3 Antibody.
Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with a cDNA expression construct encoding full-length human TRAF6 (+), using TRAF6 (D21G3) Rabbit mAb.
Confocal immunofluorescent analysis of HT-1080 (upper) and HEK/293 (lower) cells, untreated (left) or treated with TNF-α #8902 (right), using TRAF1 (45D3) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Western blot analysis of extracts from various cell lines using TRAF3 Antibody.
Western blot analysis of extracts from various cell lines using TRAF6 (D21G3) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human tonsil using TRAF1 (45D3) Rabbit mAb preincubated with a control peptide (left) or TRAF1 Blocking Peptide #1066 (right).
Western blot analysis of extracts from Raji and SR cells, using TRAF1 (45D3) Rabbit mAb.
|TRAF1 (45D3) Rabbit mAb 4715||20 µl||
|TRAF2 (C192) Antibody 4724||20 µl||
||H M Mk||53||Rabbit|
|TRAF3 Antibody 4729||20 µl||
||H M R Mk||62||Rabbit|
|TRAF6 (D21G3) Rabbit mAb 8028||20 µl||
||H Mk||60||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
The TRAF Antibody Sampler Kit provides an economical means to evaluate endogenous levels of TRAF1, 2, 3, and 6. The kit contains enough primary and secondary antibodies to perform two western mini-blot experiments.
TRAF1 (45D3) Rabbit mAb detects endogenous levels of total TRAF1 protein. TRAF2 (C192) Antibody detects endogenous levels of total human TRAF2 protein. TRAF3 Antibody detects endogenous levels of total TRAF3 protein. TRAF6 (D21G3) Rabbit mAb detects endogenous levels of total TRAF6 protein. Cross-reactivity was not detected with other family members at physiological conditions.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Cys57 of human TRAF1 or near the amino terminus of human TRAF6. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Cys192 of human TRAF2 or in the central region within TRAF3. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
TRAFs (TNF receptor-associated factors) are a family of multifunctional adaptor proteins that bind to surface receptors and recruit additional proteins to form multiprotein signaling complexes capable of promoting cellular responses (1-3). Members of the TRAF family share a common carboxy-terminal "TRAF domain", which mediates interactions with associated proteins; many also contain amino-terminal Zinc/RING finger motifs. The first TRAFs identified, TRAF1 and TRAF2, were found by virtue of their interactions with the cytoplasmic domain of TNF-receptor 2 (TNFRII) (4). The six known TRAFs (TRAF1-6) act as adaptor proteins for a wide range of cell surface receptors and participate in the regulation of cell survival, proliferation, differentiation, and stress responses.
While TRAF2 was originally described through its interaction with TNFRII, it has since been shown to interact with other surface receptors including CD27, CD30, CD40, 4-1BB, Ox40, HVEM/ATAR, and LMP-1 (1-3). TRAF2 also associates with a large number of intracellular proteins, including TRADD, FADD, I-TRAF/TANK, TRIP, A20, c-IAP1 and 2, Casper, RIP, and NIK, which help to regulate cell survival. Dominant negative and knockout studies have shown that TRAF2 plays an important role in TNF-mediated activation of NF-κB and the MAPK/JNK kinase pathway (5-7).
TRAF6 plays a critical role in innate and adaptive immunity, bone metabolism, and development of certain tissues including the nervous system (8). TRAF6 deficiency results in osteopetrosis and defective IL-1, CD40, and LPS signaling (9) as well as defects in neuronal development (10). Unlike other TRAF family members that mediate signaling through TNF, TRAF6 has unique binding activities (11) that result in signaling responses from the interleukin-1 receptor (IL-1R) (12), toll-like receptor (13,14), CD (15), RANK (16,17), and p75 neurotrophin receptor (18). TRAF6 associates directly with CD40 and RANK, and indirectly with IL-1R/TLR through IRAK (13). This leads to activation of NF-κB and MAP kinase signaling pathways through downstream association with the TAB/TAK-1 complex (19). TRAF6 also activates Src family nonreceptor tyrosine kinases leading to Akt activation (20).
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