TRAF3 Antibody (Rabbit mAb) [E9G8]

Application: Reactivity: Mouse, Rat, Human

Usage Information

Dilution
WB1:1000

Application
WB

Reactivity
Mouse, Rat, Human

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
64 kDa 64 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Positive Control	Mouse brain tissue; Rat spleen tissue; Rat brain tissue; HAP1 cells; Daudi cells; HEK-293T cells; C6 cells; RAW264.7 cells; PC-12 cells; NIH/3T3 cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
TRAF3 Antibody (Rabbit mAb) [E9G8] detects endogenous levels of total TRAF3 protein.

Clone
E9G8

Synonym(s)
CAP-1, CRAF1, TRAFAMN, TRAF3, TNF receptor-associated factor 3, CD40 receptor-associated factor 1, CD40-binding protein, LMP1-associated protein 1, RING-type E3 ubiquitin transferase TRAF3, CD40BP, LAP1

Background
TRAF3 belongs to the TNF receptor-associated factor family of cytoplasmic adaptor proteins that couple TNF receptor superfamily members to downstream signaling cascades, and it functions as a central negative regulator of the noncanonical NF-κB pathway, a signaling branch distinct from canonical NF-κB activation in that it relies on inducible processing of the precursor protein p100 rather than degradation of IκBα. Under unstimulated conditions, newly synthesized NF-κB-inducing kinase, NIK, is immediately bound by TRAF3, which recruits it into a multi-subunit E3 ubiquitin ligase complex assembled through TRAF3 dimerization with TRAF2 and the associated ubiquitin ligases cIAP1 and cIAP2; within this complex, cIAP1/2 catalyze K48-linked ubiquitination of NIK, targeting it for continuous proteasomal degradation and keeping noncanonical NF-κB signaling switched off in resting cells. Receptor engagement by ligands such as CD40 or BAFF recruits TRAF2, TRAF3, and cIAP1/2 to the receptor complex, where TRAF2 becomes activated, likely through receptor-induced aggregation, and mediates K63-linked ubiquitination of cIAP1/2, a modification that stimulates the K48-specific ligase activity of cIAP1/2 toward TRAF3 itself rather than toward NIK. This redirected ligase activity results in K48-linked ubiquitination and proteasomal degradation of TRAF3, and because TRAF3 is required to deliver NIK to the destruction complex, its loss allows newly synthesized NIK to escape degradation and accumulate, at which point NIK phosphorylates IKKα, which in turn phosphorylates p100, triggering its ubiquitination, partial proteasomal processing to p52, and nuclear translocation of p52-RelB NF-κB dimers. TRAF2 and TRAF3 play nonredundant, complementary roles within this ubiquitination cascade, with TRAF3 serving as the direct NIK-recruiting adaptor and TRAF2 serving as the bridge that couples TRAF3 to the cIAP ligases, while cIAP1 and cIAP2 function redundantly with each other, since simultaneous inhibition of both is required to trigger noncanonical NF-κB activation. TRAF3 deficiency in mice is lethal, and this lethality can be rescued by concurrent loss of NIK, directly demonstrating that unrestrained NIK accumulation, rather than loss of TRAF3 function generally, drives the pathological consequence of TRAF3 loss. Genetic deficiencies affecting the TRAF3-TRAF2-cIAP destruction complex, or amplification of the NIK gene itself, are associated with aberrant noncanonical NF-κB activation and B-cell malignancies, particularly multiple myeloma, positioning TRAF3 as a defined checkpoint whose loss of function in B lymphocytes converts a tightly restrained kinase into a constitutively active driver of NF-κB-dependent B-cell survival and proliferation signaling.

Background

TRAF3 belongs to the TNF receptor-associated factor family of cytoplasmic adaptor proteins that couple TNF receptor superfamily members to downstream signaling cascades, and it functions as a central negative regulator of the noncanonical NF-κB pathway, a signaling branch distinct from canonical NF-κB activation in that it relies on inducible processing of the precursor protein p100 rather than degradation of IκBα. Under unstimulated conditions, newly synthesized NF-κB-inducing kinase, NIK, is immediately bound by TRAF3, which recruits it into a multi-subunit E3 ubiquitin ligase complex assembled through TRAF3 dimerization with TRAF2 and the associated ubiquitin ligases cIAP1 and cIAP2; within this complex, cIAP1/2 catalyze K48-linked ubiquitination of NIK, targeting it for continuous proteasomal degradation and keeping noncanonical NF-κB signaling switched off in resting cells. Receptor engagement by ligands such as CD40 or BAFF recruits TRAF2, TRAF3, and cIAP1/2 to the receptor complex, where TRAF2 becomes activated, likely through receptor-induced aggregation, and mediates K63-linked ubiquitination of cIAP1/2, a modification that stimulates the K48-specific ligase activity of cIAP1/2 toward TRAF3 itself rather than toward NIK. This redirected ligase activity results in K48-linked ubiquitination and proteasomal degradation of TRAF3, and because TRAF3 is required to deliver NIK to the destruction complex, its loss allows newly synthesized NIK to escape degradation and accumulate, at which point NIK phosphorylates IKKα, which in turn phosphorylates p100, triggering its ubiquitination, partial proteasomal processing to p52, and nuclear translocation of p52-RelB NF-κB dimers. TRAF2 and TRAF3 play nonredundant, complementary roles within this ubiquitination cascade, with TRAF3 serving as the direct NIK-recruiting adaptor and TRAF2 serving as the bridge that couples TRAF3 to the cIAP ligases, while cIAP1 and cIAP2 function redundantly with each other, since simultaneous inhibition of both is required to trigger noncanonical NF-κB activation. TRAF3 deficiency in mice is lethal, and this lethality can be rescued by concurrent loss of NIK, directly demonstrating that unrestrained NIK accumulation, rather than loss of TRAF3 function generally, drives the pathological consequence of TRAF3 loss. Genetic deficiencies affecting the TRAF3-TRAF2-cIAP destruction complex, or amplification of the NIK gene itself, are associated with aberrant noncanonical NF-κB activation and B-cell malignancies, particularly multiple myeloma, positioning TRAF3 as a defined checkpoint whose loss of function in B lymphocytes converts a tightly restrained kinase into a constitutively active driver of NF-κB-dependent B-cell survival and proliferation signaling.

References
https://pubmed.ncbi.nlm.nih.gov/22435551/ https://pubmed.ncbi.nlm.nih.gov/18997794/

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%