Western blot analysis of extracts from mouse brain, human cerebellum and rat brain tissue using PLP1 antibody (upper) and β-tubulin (D2N5G) Rabbit mAb #15115 (lower). The upper band corresponds to the full-length PLP1 while the lower band is an alternative splice form of PLP1 known as DM20.
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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
PLP1 Antibody recognizes endogenous levels of total PLP1 protein.Species Reactivity:
Human, Mouse, RatSpecies predicted to react based on 100% sequence homology:
Pig, Guinea Pig
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human PLP1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Myelin proteolipid protein (PLP1) corresponds to the majority of myelin proteins in the CNS, providing support to axons and modulating the axonal growth (1). DM20 is the result of the alternative splicing of the plp1 gene, which is linked to oligodendrocyte differentiation and survival. The imbalance of PLP1/DM20 is linked to Pelizaeus-Merzbacher disease (2,3), including mitochondrial damage (4). In addition, PLP1, but not DM20, can enter the mitochondria and participate in the metabolism of cells (5).
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