Western blot analysis of extracts from various tissues using Alpha-internexin Antibody (upper), and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
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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
Alpha-internexin Antibody recognizes endogenous levels of total Alpha-internexin protein.Species Reactivity:
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human Alha-internexin protein. Antibodies are purified by peptide affinity chromatography.
Alpha-internexin is a class-IV neuronal intermediate filament that is involved in morphogenesis of neurons and is localized mostly in synaptic vesicles, primarily found in the post-synaptic compartment where it modulates neurotransmission function by interacting with different neurotransmitter receptors (1,2). Alpha-internexin interacts with tubulin and actin, suggesting its role in axonal transport and stabilization of dendrites (2-4). Expression of Alpha-internexin occurs in embryonic stages in rat at E10 in the cortex, auditory ganglion, olfactory epithelial, spinal cord and brainstem, and expression also occurs in postnatal stages in the cerebellum in mice and humans (3). Alpha-internexin expression is altered in different neurological diseases and disorders; where it decreases in bipolar disorder and increases in schizophrenia and Alzheimer's disease (4-6). Some of these findings were generated by proteomic analysis from Alzheimer's disease and control brains (4). The overexpression of Alpha-internexin enhances the neurite outgrowth during neuronal growth factor induction (7), induces the activation caspase-3, which triggers apoptosis and eventually neuronal death (8). Alpha-internexin could be playing a role in drug addiction, where chronic cocaine exposure decreases the levels of the protein, but not with morphine exposure, suggesting the possibility to interact with different receptors (5,9).
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