Western blot analysis of extracts from rat, mouse, and human brain using LINGO-1 Antibody.
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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.
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
LINGO-1 Antibody recognizes endogenous levels of total LINGO-1 protein.Species Reactivity:
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu400 of human LINGO-1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Leucine-rich repeat and immunoglobulin domain-containing protein (LINGO-1) is a potent negative modulator of neuronal processes including neuronal survival, axonal integrity, oligodendrocyte differentiation, and myelination (1-5). LINGO-1, Nogo receptor (NgR), and p75 neurotrophin receptor (p75NTR), or TNF receptor orphan Y (TROY) form a tripartite receptor complex, which activates RhoA/ROCK signaling and is responsible for the inhibition effect of myelin- associated factors (6,7). LINGO-1 is abundantly expressed in the brain and is implicated in various neurodegenerative disorders such as Essential tremor, multiple sclerosis and Parkinson’s disease (8-11). Recently, LINGO-1 was reported to bind directly to amyloid precursor protein (APP), promoting its degradation through lysosomal proteolysis (12). This research study implicated that Lingo-1 plays a critical role in the pathophysiology of Alzheimer's disease.
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