Western blot analysis of extracts from mouse stomach and human heart using β1-Adrenergic Receptor Antibody.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human β1-Adrenergic Receptor (β1AR; +) or full-length human β2-Adrenergic Receptor (β2AR; +), using β1-Adrenergic Receptor Antibody (upper) or β2-Adrenergic Receptor (D6H2) Rabbit mAb #8513 (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
β1-Adrenergic Receptor Antibody recognizes endogenous levels of total human β1-Adrenergic Receptor protein. This antibody does not cross-react with β2-Adrenergic Receptor.Species Reactivity:
Human, Mouse, RatSpecies predicted to react based on 100% sequence homology:
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly406 of human β1-Adrenergic Receptor protein. Antibodies are purified by protein A and peptide affinity chromatography.
β1-Adrenergic Receptor (β1AR) is a G protein-coupled receptor (GPCR) involved in the regulation of cardiovascular functions (1). Together with β2AR, β1AR is a major βAR in the heart. β1AR is activated by catecholamines and couples to Gαs protein, activating adenylate cyclase and increasing intracellular cAMP levels (2). Beta-blockers (βAR antagonists), one of the major class of therapeutics in cardiovascular medicine, act mostly by preventing catecholamine binding to β1AR (3).
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