|H M R||Endogenous||40||Rabbit|
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.
Stargazin Antibody detects endogenous levels of total stargazin protein.
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues of human stargazin. Antibodies are purified by protein A and peptide affinity chromatography.
Stargazin is a four-pass transmembrane protein related to VDCC (voltage dependent calcium channel) γ subunits and part of the TARP (transmembrane AMPA receptor regulatory protein) family of proteins. TARP proteins can form a complex with AMPA receptors (GluR1-4) and serve as integral auxiliary subunits (1-6).
Interactions between stargazin and AMPA receptors are implicated in regulation of receptor surface expression, synaptic clustering and recycling, as well as increased receptor responsiveness to glutamate (1,2,5,6). Stargazin may play a role in the molecular mechanism of AMPAR-mediated inflammatory pain by taking part in signaling pathways that relay pain in the spinal cord (5). Because the protein also modulates the pharmacology of AMPA receptors, it enhances the effects of AMPAR potentiators that have therapeutic potential for a number of mental and neurodegenerative diseases (6).
The carboxy terminus of the stargazin protein interacts with the PDZ domains of PSD95 and other membrane-associated guanylate kinase (MAGUK) family members, and together traffic AMPA receptors to the cell surface membrane, anchoring them to the postsynaptic site (1,7). Phosphorylation of stargazin by PKA on Thr321 inhibits this binding (3).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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|2503S||100 µl||$ 255.0|