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20α hydroxycholesterol

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20α-hydroxycholesterol is a chemical compound used in laboratory research. It is a naturally occurring oxysterol derived from cholesterol. The core function of 20α-hydroxycholesterol is to serve as a research tool for investigating biological processes and pathways related to cholesterol metabolism and signaling.

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Lab products found in correlation

Lovastatin, by Merck Group (2 mentions) 22 s hydroxycholesterol, by Merck Group (2 mentions) Simvastatin, by Merck Group (1 mentions) 7β hydroxycholesterol, by Merck Group (1 mentions) Gw3965, by Cayman Chemical (1 mentions) Antibodies for flow cytometry, by Thermo Fisher Scientific (1 mentions) 22 r hydroxycholesterol, by Merck Group (1 mentions) 5α cholest 7 en 3β ol, by Merck Group (1 mentions) Overexpression plasmids, by Addgene (1 mentions) 25 hydroxycholesterol, by Merck Group (1 mentions) β cyclodextrin, by Merck Group (1 mentions) 5α cholestane, by Merck Group (1 mentions) Xav939, by Selleck Chemicals (1 mentions) Igf 1, by Proteintech (1 mentions) 5 azacytidine, by Selleck Chemicals (1 mentions) Lithium chloride (licl), by Merck Group (1 mentions) Forskolin, by Selleck Chemicals (1 mentions) Sb431542, by Selleck Chemicals (1 mentions) Potassium hydroxide, by Thermo Fisher Scientific (1 mentions) Sodium chloride (nacl), by Avantor (1 mentions)

4 protocols using 20α hydroxycholesterol

1

Cholesterol Metabolism Pathway Assays

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Antibodies for flow cytometry were purchased from eBioscience. Antibodies for Western blot against Srebp1, Srebp2, LXRs, and Sumo1, -2, and -3 were purchased from Abcam, and other Western blot antibodies were purchased from Cell Signaling Technology. Cytokines were purchased from R&D Systems, and neutralization antibodies were purchased from Bio X Cell unless otherwise indicated. Cholesterol, 24(S)-hydroxycholesterol, 25-hydroxycholesterol, 7β-hydroxycholesterol, 20α-hydroxycholesterol, 3β-Hydroxy-5-cholesten-7-one, 22(R)-hydroxycholesterol, 22(S)-hydroxycholesterol, 5β-Cholestan-3β-ol, 5α-Cholest-7-en-3β-ol, 5,24-Cholestadien-3β-ol, 5-α-cholestane, β-cyclodextrin, simvastatin, lovastatin, and 2-D08 were purchased from Sigma-Aldrich. T0 and GW3965 were purchased from Cayman Chemical. Overexpression plasmids were purchased from Addgene unless otherwise indicated. All experiments were conducted according to the manufacturer’s protocols unless otherwise indicated.
Ma X., Bi E., Huang C., Lu Y., Xue G., Guo X., Wang A., Yang M., Qian J., Dong C, & Yi Q. (2018). Cholesterol negatively regulates IL-9–producing CD8+ T cell differentiation and antitumor activity. The Journal of Experimental Medicine, 215(6), 1555-1569.
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2

Induction of Cellular Reprogramming

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Small molecules were dissolved according to the manufacturer's instructions and were then processed for use at the following final concentrations: PDGFAA (Proteintech), 10 ng/mL; activin B (Proteintech), 2 μM; DAPT (Selleck), 1 μM; XAV939 (Selleck), 20 μM; forskolin (Selleck), 10 ng/mL; LH (Sigma), 1 μg/mL; 20α-hydroxycholesterol (Sigma), 10 ng/mL; IGF1 (Proteintech) 50 ng/mL; 5-azacytidine (Selleck), 5 μM; LiCl (Sigma) 5 mM; SB431542 (Selleck) 1 μM, and T3 (Sigma), 10 mM. Passaged MEFs and HPLFs (purchased from ScienCell) were seeded at a density of 100,000 cells per well in six-well plates and cultured in medium (DMEM, 10% FBS) overnight. Then, the medium was replaced with induction medium (DMEM supplemented with 10% FBS) containing the following small molecules: PDGFAA, activin B, DAPT, XAV939, forskolin, LH, 20α-hydroxycholesterol, IGF1, 5-azacytidine, LiCl, SB431542, and T3. The induction medium containing small molecules was replaced every 2 days.
Yang Y., Zhou C., Zhang T., Li Q., Mei J., Liang J., Li Z., Li H., Xiang Q., Zhang Q., Zhang L, & Huang Y. (2020). Conversion of Fibroblast into Functional Leydig-like Cell Using Defined Small Molecules. Stem Cell Reports, 15(2), 408-423.
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3

Cholesterol Oxidation Compounds Analysis

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The chemicals and reagents were all of analytical grade (Sigma grade ≥95% purity). The cholesterol and cholesterol oxide standards of cholesterol (5-cholesten-3β-ol), 5α-cholestane, 6-ketocholesterol (5α-cholestan-3β-ol-6-one), α-epoxide (5α, 6α-epoxycholestan-3β-ol), β-epoxide (5β, 6β-epoxycholestan-3β-ol) and 20α-hydroxycholesterol were from Sigma Aldrich Inc. (St Louis, MO, USA). Potassium hydroxide was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Butylated hydroxytoluene (BHT), Celite 545, dicalcium phosphate dehydrate (CAHPO4 × 2H2O), pyridine, BSTFA (N,O-bis(trismethylsilyl) trifluoroacetamide) and TMCS (trimethylchlorosilane) (BSTFA + 1% TMCS), and 1,4-dioxane were also purchased from Sigma Aldrich Inc. (St Louis, MO, USA). Methanol and diethyl ether were from Riedel-de Haen, Seelze, Germany. Trichloromethane, sulphuric acid, fydrochloric acid (32%), hexane, ethyl acetate, and acetone were from Fisher Scientific (Poole, England). Sodium chloride was from BDH Chemicals (Poole, England).
Mungure T.E., Birch E.J., Ponnampalam E.N., Stewart I., Ahmed I.A., Al-Juhaimi F.Y, & Bekhit A.E. (2020). Conjugated Linoleic Acid and Cholesterol Oxidative Products Generated in Hot Boned Beef Semimembranosus Muscle as Affected by Rigor Temperature, Ageing and Display Time. Foods, 9(1), 43.
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4

Cholesterol Metabolite Modulation of Shh Signaling

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Water soluble SAG was purchased from Calbiochem. Shh-N was produced in bacteria according to (88 (link)). BM15.766 (Santa Cruz) and AY9944 (Calbiochem) were solubilized in Dimethyl sulfoxide (DMSO). BM15.766 was present at a concentration of 15 μm and AY9944 at 10 μm in assays unless stated otherwise. Lovastatin (Sigma) solubilized in H2O was present at a concentration of 5 μm in assays. 7DHC, 22(S)-hydroxyCholesterol and 20α-hydroxyCholesterol (all Sigma) were solubilized in ethanol. Cholesterol (Sigma) was solubilized in 30% 2-hydroxypropyl-β-cyclodextrin (Sigma). Cholesterol and 7DHC were present at 50 μm, and 22S- and 20α-hydroxyCholesterol were present at 10 μm in assays. In all cases equivalent concentrations of vehicle were added to controls. When present 2-hydroxypropyl-β-cyclodextrin was at a concentration of 0.1%.
Blassberg R., Macrae J.I., Briscoe J, & Jacob J. (2015). Reduced cholesterol levels impair Smoothened activation in Smith–Lemli–Opitz syndrome. Human Molecular Genetics, 25(4), 693-705.
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