|H M||Endogenous||50||Rabbit IgG|
Western blot analysis of extracts from various cell lines using TRAF4 (D1N3A) Rabbit mAb.Learn more about how we get our images
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human TRAF4 (hTRAF4-Myc/DDK; +), using TRAF4 (D1N3A) Rabbit mAb (upper) or Myc-Tag (71D10) Rabbit mAb #2278 (lower).Learn more about how we get our images
Western blot analysis of extracts from SK-BR-3 cells, mock transfected (-) or transfected with siRNA against human TRAF4, using TRAF4 (D1N3A) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).Learn more about how we get our images
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised October 2016
Protocol Id: 10
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
TRAF4 (D1N3A) Rabbit mAb recognizes endogenous levels of total TRAF4 protein. An unknown background band is detected in some cell lines at 80kDa.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg124 of human TRAF4 protein.
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.
TRAF4, also referred to as CART1 and MLN62, is a divergent member of the TRAF family with relatively weak binding to TNFR family members (5-7). Interactions have been observed between TRAF4 and the neurotrophin receptor p75-NGFR, lymphotoxin-β receptor, and GITR (8-10). While originally identified in metastatic breast carcinoma, TRAF4 has been shown to contribute to tumor growth and invasion in various cancers including breast, lung and colon (11-13). Expression of Traf4 is induced by the tumor suppressor p53 in response to DNA damage, and can promote apoptosis (14).TRAF4 has also been shown to play a critical role in TGF-β signaling, where it has been found to antagonize the E3 ligase Smurf, resulting in enhanced receptor stabilization driving breast cancer metastasis (15).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|18527S||100 µl (10 western blots)||$ 255.0|