Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human 5-HTR4 (h5-HTR4-Myc/DDK; +), using 5-HTR4 (D8O5K) Rabbit mAb.
Western blot analysis of extracts from NIH/3T3 and PC-12 cells using 5-HTR4 (D8O5K) Rabbit mAb.
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Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. 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
5-HTR4 (D8O5K) Rabbit mAb recognizes endogenous levels of total 5-HTR4 protein.Species Reactivity:
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala361 of human 5-HTR4 protein.
Serotonin (5-hydroxytryptamine, 5-HT) is a central nervous system monoamine implicated in a number of physiological functions, including sleep, mood, appetite, anxiety, and neurovegetative control (1). Serotonin acts through the activation of G protein receptors called serotonin receptors (5-HTRs). Seven 5-HTR subclasses (5-HTR1-7) have been described to date (2). Serotonin receptor 4 (5-HTR4) couples to Gαs (3), with stimulation of the receptor leading to adenylyl cyclase activation, elevation of cyclic AMP levels and downstream PKA activation (4). The corresponding HTR4 gene exhibits a complex structure, which includes a large number of splice variants that seem to share the same pharmacological properties (5). 5-HTR4 is one of several receptors implicated in cognition, depression, and the development of Alzheimer's disease (6-8).
Explore pathways + proteins related to this product.
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