Western blot analysis of human umbilical vein endothelial cells, untreated (-) or treated with TNF-α #8902 (50 ng/ml, 24 hr; +), using TNFSF15 (D1C8W) Rabbit mAb (upper) and β-Actin (13E5) Rabbit mAb #4970 (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 10mM sodium HEPES (pH 7.5), 150mM NaCl, 100 μg/ml, 50% glycerol and less than 0.02% sodium azide. Store at -20ºC. Do not aliquot the antibody.
TNFSF15 (D1C8W) Rabbit mAb recognizes endogenous levels of total human TNFS15 protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu88 of human TNFSF15 protein.
TNFSF15 (TL1A), a member of the TNF superfamily of proteins, is a splice variant of the TL1/VEGI gene (1). Endothelial cells, monocytes, macrophages, and dendritic cells express TL1A, which is upregulated by proinflammatory cytokines, microorganisms, and AMPK activation (1-4). TL1A activates the NF-κB and JNK pathways through its receptor, DR3 (1,5). TL1A may function as a costimulatory signal for T cell activation, specifically regulating Th17 cell development and proliferation (1,2,6). Mouse models suggest a role for TL1A as a driver for the inflammation and pathogenesis associated with inflammatory bowel disease (7,8).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|57221S||100 µl (10 western blots)||$ 255.0|