This antibody gave a positive signal in the following tissue lysates: Human lung; Human kidney; Human heart; Human placenta; Human skeletal muscle as well as the following whole cell lysates: HeLa; Jurkat; K562; PC3; NIH3T3.
存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa).
Use a concentration of 1 µg/ml.
Use a concentration of 5 µg/ml.
功能E3 ubiquitin ligase that, together with UBE2N and UBE2V1, mediates the synthesis of 'Lys-63'-linked-polyubiquitin chains conjugated to proteins, such as IKBKG, AKT1 and AKT2. Also mediates ubiquitination of free/unanchored polyubiquitin chain that leads to MAP3K7 activation. Leads to the activation of NF-kappa-B and JUN. May be essential for the formation of functional osteoclasts. Seems to also play a role in dendritic cells (DCs) maturation and/or activation. Represses c-Myb-mediated transactivation, in B lymphocytes. Adapter protein that seems to play a role in signal transduction initiated via TNF receptor, IL-1 receptor and IL-17 receptor.
组织特异性Expressed in heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
通路Protein modification; protein ubiquitination.
序列相似性Belongs to the TNF receptor-associated factor family. A subfamily. Contains 1 MATH domain. Contains 1 RING-type zinc finger. Contains 2 TRAF-type zinc fingers.
结构域The coiled coil domain mediates homo- and hetero-oligomerization. The MATH/TRAF domain binds to receptor cytoplasmic domains.
翻译后修饰Sumoylated on Lys-124, Lys-142 and Lys-453 by SUMO1. Polyubiquitinated on Lys-124; after cell stimulation with IL-1-beta or TGF-beta. This ligand-induced cell stimulation leads to dimerization/oligomerization of TRAF6 molecules, followed by auto-ubiquitination which involves UBE2N and UBE2V1 and leads to TRAF6 activation. This 'Lys-63' site-specific poly-ubiquitination appears to be associated with the activation of signaling molecules. Endogenous autoubiquitination occurs only for the cytoplasmic form.
细胞定位Cytoplasm. Cytoplasm > cell cortex. Nucleus. Found in the nuclei of some agressive B-cell lymphoma cell lines as well as in the nuclei of both resting and activated T-and B-lymphocytes. Found in punctate nuclear body protein complexes. Ubiquitination may occur in the cytoplasm and sumoylation in the nucleus.
TRAF6 was immunoprecipitated using 0.5mg Mouse Skeletal Muscle tissue lysate, 5µg of Rabbit polyclonal to TRAF6 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Skeletal Muscle tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab94720.
All lanes : Anti-TRAF6 antibody (ab94720) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 3 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate Lane 4 : PC3 (Human prostate carcinoma cell line) Whole Cell Lysate Whole Cell Lysate Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 6 : Lung (Human) Tissue Lysate Lane 7 : Human kidney tissue lysate - total protein (ab30203) Lane 8 : Human heart tissue lysate - total protein (ab29431) Lane 9 : Human placenta tissue lysate - total protein (ab29745) Lane 10 : Human skeletal muscle tissue lysate - total protein (ab29330)
Lysates/proteins at 10 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 60 kDa Observed band size : 60 kDa Additional bands at : 100 kDa. We are unsure as to the identity of these extra bands.
ICC/IF image of ab94720 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab94720 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in formaldehyde fixed (4%, 10min) MCF-7 cell types at 1ug/ml
Anti-TRAF6 antibody (ab94720)参考文献
has not yet been referenced specifically in any publications.