For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
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.
Do not add Anti-biotin, HRP-linked Antibody for detection of biotinylated protein markers. There is no need. The Streptavidin-HRP will also visualize the biotinylated markers.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 265
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly653 of human CD109 protein.
CD109 is a glycosylphosphatidylinositol (GPI)-linked glycoprotein that belongs to the alpha2-macroglobulin family of thioester containing proteins (1). CD109 is associated with TGF-beta receptor I (TbRI) and inhibits TGF-beta signaling (2,3). Cleavage of CD109 at its Furin cleavage site results in the release of its large amino-terminal domain, which then binds to the TGF-beta receptor I to inhibit TGF-beta signaling (4-7). CD109 is expressed on a subset of CD34+ bone marrow cells and mesenchymal stem cells, activated platelets, activated T cells, endothelial cells, and a wide variety of tumors (8-10). Elevated CD109 expression has been considered a diagnostic/prognostic marker for several types of cancers (11-14).
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