25-Hydroxycholesterol is a major metabolite of cholesterol produced by macrophages in response to the activation of Toll-like receptor (TLR). 25-hydroxycholesterol is a potent (EC50≈65 nM) and selective suppressor of IgA production by B cells.

Physicochemical Properties

Molecular Formula	C27H46O2
Molecular Weight	402.6529
Exact Mass	402.349
CAS #	2140-46-7
PubChem CID	65094
Appearance	White to off-white solid powder
Density	1.0±0.1 g/cm3
Boiling Point	513.1±23.0 °C at 760 mmHg
Flash Point	213.5±17.2 °C
Vapour Pressure	0.0±3.0 mmHg at 25°C
Index of Refraction	1.538
LogP	7.66
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	2
Rotatable Bond Count	5
Heavy Atom Count	29
Complexity	636
Defined Atom Stereocenter Count	8
SMILES	O([H])[C@@]1([H])C([H])([H])C([H])([H])[C@@]2(C([H])([H])[H])C(C1([H])[H])=C([H])C([H])([H])[C@]1([H])[C@]2([H])C([H])([H])C([H])([H])[C@]2(C([H])([H])[H])[C@@]([H])([C@]([H])(C([H])([H])[H])C([H])([H])C([H])([H])C([H])([H])C(C([H])([H])[H])(C([H])([H])[H])O[H])C([H])([H])C([H])([H])[C@]21[H]
InChi Key	INBGSXNNRGWLJU-ZHHJOTBYSA-N
InChi Code	InChI=1S/C27H46O2/c1-18(7-6-14-25(2,3)29)22-10-11-23-21-9-8-19-17-20(28)12-15-26(19,4)24(21)13-16-27(22,23)5/h8,18,20-24,28-29H,6-7,9-17H2,1-5H3/t18-,20+,21+,22-,23+,24+,26+,27-/m1/s1
Chemical Name	(3S,8S,9S,10R,13R,14S,17R)-17-[(2R)-6-hydroxy-6-methylheptan-2-yl]-10,13-dimethyl-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-ol
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro	The enzyme cholesterol 25-hydroxylase catalyzes the production of 25-hydroxycholesterol by using NADPH as a cofactor and cholesterol and molecular oxygen as substrates. A powerful bioactive lipid in both the innate and adaptive immune systems is 25-hydroxycholesterol[1].
References	[1]. McDonald JG, et al. Editorial: 25-Hydroxycholesterol: a new life in immunology. J Leukoc Biol. 2010 Dec;88(6):1071-2.
Additional Infomation	25-hydroxycholesterol is an oxysterol and a 25-hydroxy steroid. It has a role as a human metabolite. It is functionally related to a cholesterol. 25-Hydroxycholesterol has been reported in Bombyx mori, Homo sapiens, and Rissoella verruculosa with data available.

Solubility Data

Solubility (In Vitro)

Ethanol : ~14.29 mg/mL (~35.49 mM) DMSO : ~5 mg/mL (~12.42 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 0.5 mg/mL (1.24 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 5.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 2: ≥ 0.5 mg/mL (1.24 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 5.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 3: 10 mg/mL (24.84 mM) in 50% PEG300 50% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.4835 mL	12.4177 mL	24.8355 mL
	5 mM	0.4967 mL	2.4835 mL	4.9671 mL
	10 mM	0.2484 mL	1.2418 mL	2.4835 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.