Western blot analysis of extracts from various cell lines using LXR-β (D6M9D) Rabbit mAb.
Western blot analysis of extracts from 293 cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human LXR-α protein (hLXR-α-Myc/DDK; +) or Myc/DDK-tagged full-length human LXR-β protein (hLXR-β-Myc/DDK; +), using LXR-β (D6M9D) Rabbit mAb (upper), DYKDDDDK (9A3) Mouse mAb #8146 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
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
LXR-β (D6M9D) Rabbit mAb recognizes endogenous levels of total LXR-β protein.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human LXR-β protein.
Liver X receptors LXR-α and LXR-β are nuclear hormone receptor superfamily members responsible for regulating expression of target genes that control cholesterol transport and metabolism (1). When bound by the oxidized derivatives of cholesterol (oxysterols), activated LXR receptors function as sterol sensors to regulate transcription of the genes involved in the cholesterol homeostasis (1,2). The LXR-α protein is expressed at high levels in rat liver, kidney, intestine, adipose, and spleen; LXR-β is more ubiquitously expressed within rat tissues (1,3). Research studies indicate that glucose binds and up-regulates the transcriptional activity of LXR-α and LXR-β (4). LXR-α and LXR-β are putative glucose sensors that integrate glucose metabolism and fatty acid biosynthesis in the liver (4). Additional studies show that female mice deficient in LXR-β develop gallbladder cancer (5). In addition, LXR-β plays a role in protecting dopaminergic neurons in a Parkinson disease model (6).
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