GLPBio

20(S)-Hydroxycholesterol

Catalog No.: GC15422Purity:>98.00%Synonyms: 20α-hydroxy Cholesterol
COASDSData Sheet
20(S)-Hydroxycholesterol is a selective allosteric activator of the Hedgehog pathway protein Smoothened (EC50=3μM).

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20(S)-Hydroxycholesterol
Cas No.: 516-72-3
10mM (in 1mL DMSO)In Stock
$131.00
1
1mgIn Stock
$50.00
1
5mgIn Stock
$118.00
1
10mgIn Stock
$191.00
1
25mgIn Stock
$323.00
1
50mgIn Stock
$467.00
1
100mgIn Stock
$663.00
1
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Description

20(S)-Hydroxycholesterol is a selective allosteric activator of the Hedgehog pathway protein Smoothened (EC50=3μM). 20(S)-Hydroxycholesterol binds to the extracellular cysteine-rich domain (CRD) of Smoothened in a stereoselective manner, activating downstream Gli transcription factors. 20(S)-Hydroxycholesterol also enhances osteogenic differentiation of bone marrow stromal cells and synergizes with simvastatin to activate the Raf/MEK/ERK pathway, promoting bone regeneration. 20(S)-Hydroxycholesterol can be used in research related to developmental biology, oncology, and bone and angiogenesis mechanisms[1-4].

In vitro, 20(S)-Hydroxycholesterol (5μM) treatment of cortical neurons cultured on reactive astrocytes for 24 hours significantly increased neurite outgrowth. Treatment of cortical neurons with 20(S)-Hydroxycholesterol (5μM) for 30 minutes significantly increased Y416-Src phosphorylation[5]. A combination of 20(S)-hydroxycholesterol and 22(S)-hydroxycholesterol (1:1) (5μM) was used to treat M2-10B4 mouse bone marrow stromal cells for 10 minutes to 8 hours, followed by culture in differentiation medium for 4 to 14 days. This treatment induced alkaline phosphatase activity, increased mineralization, elevated osteocalcin mRNA expression, and promoted nuclear protein binding to the OSE2 site of the osteocalcin promoter[6].

References:
[1] Nachtergaele S, Mydock LK, Krishnan K, et al. Oxysterols are allosteric activators of the oncoprotein Smoothened. Nat Chem Biol. 2012 Jan 8;8(2):211-20.
[2] Nedelcu D, Liu J, Xu Y, et al. Oxysterol binding to the extracellular domain of Smoothened in Hedgehog signaling. Nat Chem Biol. 2013 Sep;9(9):557-64.
[3] Cheng YS, Zhang T, Ma X, et al. A proteome-wide map of 20(S)-hydroxycholesterol interactors in cell membranes. Nat Chem Biol. 2021 Dec;17(12):1271-1280.
[4] Peyrot SM, Nachtergaele S, Luchetti G, et al. Tracking the subcellular fate of 20(s)-hydroxycholesterol with click chemistry reveals a transport pathway to the Golgi. J Biol Chem. 2014 Apr 18;289(16):11095-11110.
[5] Berretta A, Gowing EK, Jasoni CL, et al. Sonic hedgehog stimulates neurite outgrowth in a mechanical stretch model of reactive-astrogliosis. Sci Rep. 2016 Feb 23;6:21896.
[6] Aghaloo TL, Amantea CM, Cowan CM, et al. Oxysterols enhance osteoblast differentiation in vitro and bone healing in vivo. J Orthop Res. 2007 Nov;25(11):1488-97.

Protocol

Cell experiment [1]:

Cell lines

M2-10B4 mouse marrow stromal cells

Preparation Method

M2-10B4 cells were cultured in RPMI supplemented with 5% fetal bovine serum (FBS), 50μg/mL ascorbic acid and 3mM β-glycerophosphate. Cells were treated with 5μM of the osteogenic oxysterol cocktail 20(S)-hydroxycholesterol (20S) + 22(S)-hydroxycholesterol (22S) (SS, 1:1) for indicated time periods (10min to 8h), then washed twice and cultured in differentiation medium for the duration indicated for each experiment.

Reaction Conditions

5μM 20(S)-hydroxycholesterol; 10min to 8h

Applications

20(S)-hydroxycholesterol treatment for 10min significantly increased alkaline phosphatase (ALP) activity after 4 days. 20(S)-hydroxycholesterol treatment for 2h significantly increased mineralization after 14 days. 20(S)-hydroxycholesterol treatment for 2-8h increased osteocalcin (OCN) mRNA expression measured after 8 days. 20(S)-hydroxycholesterol treatment for 0.5-24h induced nuclear protein binding to an OSE2 site measured after 4 days.

References:
[1] Aghaloo TL, Amantea CM, Cowan CM, et al. Oxysterols enhance osteoblast differentiation in vitro and bone healing in vivo. J Orthop Res. 2007 Nov;25(11):1488-97.

Product Documents

Purity:>98.00%Appearance:A solid
COASDSData Sheet

Chemical Properties

CAS No.
516-72-3
Synonyms
20α-hydroxy Cholesterol
Chemical Name
(3S,8S,9S,10R,13S,14S,17S)-17-((S)-2-hydroxy-6-methylheptan-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol
SMILES
O[C@](CCCC(C)C)(C)[C@@H]1[C@](CC2)(C)[C@@H](CC1)[C@H]3[C@H]2[C@](CC[C@@H]4O)(C)C(C4)=CC3
Molecular Formula
C27H46O2
Molecular Weight
402.65
Solubility
Ethanol:50mg/ml
Storage
Store at -20°C
General tips
Select a suitable solvent according to the product solubility data. Once prepared, aliquot the stock solution to avoid repeated freeze-thaw cycles.
Use stock solutions within 6 months when stored at -80°C or within 1 month when stored at -20°C.
To improve solubility, warm the tube to 37°C and sonicate it for an appropriate period.
Shipping Condition
Evaluation samples are shipped with blue ice. Other available sizes are shipped at room temperature or with blue ice upon request.

CalculatorsMolarity / Dilution / Molecular Weight / Unit Conversion / In Vivo Formulation / Solubility

g/mol

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