25-Hydroxycholesterol (Synonyms: 25-OHC)

Name: 25-Hydroxycholesterol
Brand: MedChemExpress
SKU: HY-113134
Rating: 5.0 (28 reviews)

Cat. No.: HY-113134 Purity: 99.92%: Data Sheet
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25-Hydroxycholesterol (25-OHC) is a metabolite of cholesterol that is produced and secreted by macrophages in response to Toll-like receptor (TLR) activation. 25-hydroxycholesterol is a potent (EC₅₀≈65 nM) and selective suppressor of IgA production by B cells.

For research use only. We do not sell to patients.

CAS No. : 2140-46-7

Size	Stock	Quantity

Free Sample (0.1 - 0.2 mg)	Apply Now
Solid + Solvent (Highly Recommended)
10 mM * 1 mL in DMSO ready for reconstitution	In-stock	Estimated Time of Arrival: December 31
Solution
10 mM * 1 mL in DMSO	In-stock	Estimated Time of Arrival: December 31
Solid
5 mg	In-stock	Estimated Time of Arrival: December 31
10 mg	In-stock	Estimated Time of Arrival: December 31
25 mg	In-stock	Estimated Time of Arrival: December 31
50 mg	In-stock	Estimated Time of Arrival: December 31
100 mg	In-stock	Estimated Time of Arrival: December 31
200 mg	Get quote
500 mg	Get quote

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Based on 28 publication(s) in Google Scholar

Other Forms of 25-Hydroxycholesterol:

25-Hydroxycholesterol-d₆ In-stock
25-Hydroxycholesterol (Standard) Get quote

28 Publications Citing Use of MCE 25-Hydroxycholesterol

RT-PCR

Cell Proliferation/Viability Assay

Flow Cytometry

: 25-Hydroxycholesterol purchased from MedChemExpress. Usage Cited in: Nat Commun. 2025 Jan 31;16(1):1217. [Abstract]; 25-Hydroxycholesterol (1 µg/mL; 48 h). BMDM were incubated with B16F10-OVA VEGF-C tumor-conditioned medium (TCM), treated or not with 25-HC for 48 h, and analyzed by Q-PCR for PPAR-γ mRNA levels.

: 25-Hydroxycholesterol purchased from MedChemExpress. Usage Cited in: Nat Commun. 2025 Jan 31;16(1):1217. [Abstract]; 25-Hydroxycholesterol (1 µg/mL; 48 h). BMDMs were incubated with tumor-conditioned medium (TCM), treated or not with 25-HC, Troglitazone, or the combination of 25-HC and Troglitazone for 48 h. BMDM phenotype was analyzed by flow cytometry.

: 25-Hydroxycholesterol purchased from MedChemExpress. Usage Cited in: Chem Biol Interact. 2024 Aug 26:111214. [Abstract]; 25-Hydroxycholesterol (0-10 µM; 24-72 h). 25HC inhibits short-term proliferation in Swan 71 cells. The cells were exposed to the indicated concentrations of 25HC for 24, 48, and 72 h. Cell viability was measured using a WST assay.

: 25-Hydroxycholesterol purchased from MedChemExpress. Usage Cited in: Mol Cell. 2023 Aug 17;83(16):3010-3026.e8. [Abstract]; Co-IP analysis of LAM cell serum starved overnight (to induce SREBP cleavage) and treated with SREBP cleavage inhibitor (25-Hydroxycholesterol, 25-HC, 10 µM), or SRPK inhibitor (SRPKIN-1, 5 µM) for 4 h.

: 25-Hydroxycholesterol purchased from MedChemExpress. Usage Cited in: Cell Metab. 2022 Oct 4;34(10):1548-1560.e6. [Abstract]; HEK293 cells were transfected with the indicated plasmids and supplemented with 25-Hydroxycholesterol (25-HC, 1 μg/ml) for 16 h. Indicated proteins were IB analyzed.

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Human Endogenous Metabolite Microbial Metabolite

Biological Activity
Purity & Documentation
References
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Description

IC₅₀ & Target

Human Endogenous Metabolite

Cellular Effect

Cell Line	Type	Value	Description	References
CHO-K1	IC₅₀	0.3 μM Compound: 25-hydroxycholesterol	Inhibition of SREBP2 activation expressed in CHO-K1 cells co-transfected with pSRE-Luc plasmid assessed as inhibition of luciferase expression after 20 hrs by luciferase reporter gene assay Inhibition of SREBP2 activation expressed in CHO-K1 cells co-transfected with pSRE-Luc plasmid assessed as inhibition of luciferase expression after 20 hrs by luciferase reporter gene assay	[PMID: 21561152]
HEK293	EC₅₀	0.63 μM Compound: 25HC	Induction of human HMGCR-dCat-ELuc degradation expressed in HEK293 cells assessed as reduction in luciferase activity after 4 hrs by luciferase reporter assay Induction of human HMGCR-dCat-ELuc degradation expressed in HEK293 cells assessed as reduction in luciferase activity after 4 hrs by luciferase reporter assay	[PMID: 31902650]
HEK293	EC₅₀	2.3 μM Compound: 11, 25-HC	Binding affinity to human GFP-tagged NPC1L1 L1072T/L1168I mutant expressed in HEK293 cells assessed as localization to endoplasmic reticulum and plasma membrane after 24 hrs by fluorescence microscopic analysis Binding affinity to human GFP-tagged NPC1L1 L1072T/L1168I mutant expressed in HEK293 cells assessed as localization to endoplasmic reticulum and plasma membrane after 24 hrs by fluorescence microscopic analysis	[PMID: 23830695]
HEK293	EC₅₀	3.3 μM Compound: 1, 25HC	Binding affinity to FLAG/tGFP-tagged NPC1 I1061T mutant (unknown origin) expressed in HEK293 cells assessed as localization after 24 hrs by fluorescence microscopy Binding affinity to FLAG/tGFP-tagged NPC1 I1061T mutant (unknown origin) expressed in HEK293 cells assessed as localization after 24 hrs by fluorescence microscopy	[PMID: 24928400]
HEK293	EC₅₀	> 10000 nM Compound: 39	Positive allosteric modulation of recombinant human GluN1/GluN2A receptor stably expressed in HEK293 cells assessed as increase in glycine/L-glutamate-induced channel current at -70 mV holding potential by IonWork two-electrode voltage clamp method Positive allosteric modulation of recombinant human GluN1/GluN2A receptor stably expressed in HEK293 cells assessed as increase in glycine/L-glutamate-induced channel current at -70 mV holding potential by IonWork two-electrode voltage clamp method	[PMID: 31390523]
HEK293	EC₅₀	> 10000 nM Compound: 39	Positive allosteric modulation of recombinant human GluN1/GluN2B receptor stably expressed in HEK293 cells assessed as increase in glycine/L-glutamate-induced channel current at -70 mV holding potential by IonWork two-electrode voltage clamp method Positive allosteric modulation of recombinant human GluN1/GluN2B receptor stably expressed in HEK293 cells assessed as increase in glycine/L-glutamate-induced channel current at -70 mV holding potential by IonWork two-electrode voltage clamp method	[PMID: 31390523]
HepG2	IC₅₀	0.3 μM Compound: 25-hydroxy-cholesterol	Tested for its inhibitory activity against HMG-CoA reductase transcription in HepG2 cells Tested for its inhibitory activity against HMG-CoA reductase transcription in HepG2 cells	[PMID: 8057282]
Vero	IC₅₀	> 128 μM Compound: 7	Cytotoxicity against african green monkey Vero cells by tetrazolium dye reduction assay Cytotoxicity against african green monkey Vero cells by tetrazolium dye reduction assay	[PMID: 24084159]

In Vitro

The synthesis of 25-hydroxycholesterol is catalyzed by the enzyme cholesterol 25-hydroxylase, which uses cholesterol and molecular oxygen as substrates and NADPH as a cofactor. 25-hydroxycholesterol is a potent bioactive lipid in the innate and adaptive immune systems^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight

402.65

Formula

C₂₇H₄₆O₂

CAS No.

2140-46-7

Appearance

Solid

Color

White to off-white

SMILES

CC(C)(O)CCC[C@@H](C)[C@H]1CC[C@@]2([H])[C@]3([H])CC=C4C[C@@H](O)CC[C@]4(C)[C@@]3([H])CC[C@]12C

Structure Classification

Steroids

Initial Source

Endogenous metabolite

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	6 months
	-20°C	1 month

Solvent & Solubility

In Vitro:

Ethanol : 14.29 mg/mL (35.49 mM; Need ultrasonic)

DMSO : 5 mg/mL (12.42 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions

Concentration Solvent Mass	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

View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Molarity Calculator
Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

Concentration _(start)

Volume _(start)

Concentration _(final)

Volume _(final)

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 0.5 mg/mL (1.24 mM); Clear solution

This protocol yields a clear solution of ≥ 0.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (5.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Protocol 2

Add each solvent one by one: 10% DMSO 90% Corn Oil
Solubility: ≥ 0.5 mg/mL (1.24 mM); Clear solution

This protocol yields a clear solution of ≥ 0.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (5.0 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Please enter your animal formula composition:

DMSO +

Tween-80 +

Saline

Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.

The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).

Calculation results:

Working solution concentration: mg/mL

Method for preparing stock solution: mg drug dissolved in μL DMSO (Stock solution concentration: mg/mL).

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).

Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.

If the continuous dosing period exceeds half a month, please choose this protocol carefully.

Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.

Purity & Documentation

Purity: 99.98%

Select Batch:

Data Sheet (274 KB) SDS (422 KB)

COA (249 KB) RP-HPLC (274 KB) Elemental Analysis Report (107 KB)

Handling Instructions (2659 KB)

References

[1]. McDonald JG, et al. Editorial: 25-Hydroxycholesterol: a new life in immunology. J Leukoc Biol. 2010 Dec;88(6):1071-2. [Content Brief]

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg

DMSO / Ethanol	1 mM	2.4835 mL	12.4177 mL	24.8355 mL	62.0887 mL
	5 mM	0.4967 mL	2.4835 mL	4.9671 mL	12.4177 mL
	10 mM	0.2484 mL	1.2418 mL	2.4835 mL	6.2089 mL
Ethanol	15 mM	0.1656 mL	0.8278 mL	1.6557 mL	4.1392 mL
	20 mM	0.1242 mL	0.6209 mL	1.2418 mL	3.1044 mL
	25 mM	0.0993 mL	0.4967 mL	0.9934 mL	2.4835 mL
	30 mM	0.0828 mL	0.4139 mL	0.8278 mL	2.0696 mL

25-Hydroxycholesterol Related Classifications

Inflammation/Immunology

Help & FAQs

Do most proteins show cross-species activity?
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

25-Hydroxycholesterol2140-46-725-OHCEndogenous MetaboliteInhibitorinhibitorinhibit

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25-Hydroxycholesterol (Synonyms: 25-OHC)

Customer Review

Other Forms of 25-Hydroxycholesterol:

25-Hydroxycholesterol Related Antibodies

28 Publications Citing Use of MCE 25-Hydroxycholesterol

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•Related Screening Libraries:

•Related Small Molecules:

•Related Proteins:

View All Endogenous Metabolite Isoform Specific Products:

Biological Activity

Purity & Documentation

References

Customer Review

25-Hydroxycholesterol Related Antibodies

Molarity Calculator

Dilution Calculator

Complete Stock Solution Preparation Table

25-Hydroxycholesterol Related Classifications

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