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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 and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
BMPR2 Antibody recognizes endogenous levels of total BMPR2 protein. This antibody may also detect a 115 Kd unspecific band.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys763 of human BMPR2 protein. Antibodies are purified by protein A and peptide affinity chromatography.
BMPR2 is a type II serine/threonine receptor kinase that binds to an array of secreted bone morphogenetic proteins (BMPs). BMPs belong to the superfamily of TGF-β ligands that modulate gastrulation, neurogenesis, chondrogenesis, interdigital cell death, and bone morphogenesis (1-5). In contrast to the TGF-β type II receptor, BMPR2 contains an extended carboxyl-terminal region that interacts with multiple signaling molecules to modulate the responsiveness of target genes to BMPs (6,7). BMP signaling requires oligomerization of both type I and type II receptors to elicit a functional response of target genes. BMP binding to type I and II receptors induces Smad1/5/8 phosphorylation which is required for the activation of target genes (7). In vitro and in vivo evidence suggests that defects in BMPR2 may contribute to pulmonary hypertension, inflammation, and endothelial injury (8,9).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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