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.
Msx1 (P5) Antibody detects endogenous levels of total Msx1 protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acid sequences surrounding Pro5 of human Msx1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Msh homeobox 1 (Msx1) is a Muscle Segment Homeobox (Msh) gene family member that acts as a transcriptional repressor during embryonic development, playing an important role in limb pattern formation, craniofacial development, and tooth development (1-3). Msx1 is expressed in the mesenchyme of the developing nail bed (2) and in fetal hair follicles, epidermis and fibroblasts; reduced expression is seen in adult epithelial-derived tissues (4). Msx1 acts in concert with the Wnt1 network to establish the midbrain dopaminergic progenator domain, a region that gives rise to neurons that are critical for normal brain function and are the cells affected in Parkinson disease (5). Mutation in the corresponding Msx1 gene correlates with abnormal tooth development in patients diagnosed with Wolf-Hirschhorn syndrome (6). Other genetic changes in the Msx1 gene result in Witkop Syndrome ("tooth and nail syndrome") and cases of abnormal tooth development associated with non-syndromic orofacial clefting (2,7).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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