Western blot analysis of extracts from various cell types using Plectin-1 Antibody. Molecular weights were determined using a pre-stained molecular weight marker (not shown).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 and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
Plectin-1 Antibody detects endogenous levels of total plectin-1 protein. The antibody is expected to cross-react with all isoforms of plectin-1 based on the sequence of the immunogenic peptide.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the central sequence of human plectin-1.
Plectin is a large, widely expressed protein that crosslinks the intermediate filament and actin cytoskeleton, mechanically stabilizing cells and tissues. Plectin also plays a role in the regulation of actin dynamics and acts as a scaffold for signaling molecules (1). Plectin is important in the stabilization of hemidesmosomes, crosslinking them to the intermediate filament network. Research studies have shown that mutations in plectin and other genes coding for hemidesmosomal proteins can cause epidermolysis bullosa, a condition manifested by fragile skin and frequent blistering (1,2). Plectin modulates signals to PKC through binding and sequestration of RACK1, the receptor for activated C kinase 1 (3,4). Plectin is also involved in the regulation of cytokeratin architecture and cell stress response (4), signaling through the chemokine receptor CXCR4 (5) and regulation of AMP-activated protein kinase (AMPK) activity and signaling in mouse myotubes (6).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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|2863S||100 µl (10 western blots)||$ 255.0|