7alpha,25-OHC

Cat No.:V7622 Purity: ≥98%

COA Product Data Instructions for use SDS

7α,25-dihydroxycholesterol (7α,25-OHC) is a potent and specific agonist and endogenous ligand of the orphan GPCR receptor EBI2 (GPR183).

7alpha,25-OHC Chemical Structure CAS No.: 64907-22-8
Product category: New1

This product is for research use only, not for human use. We do not sell to patients.

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Product Description
Bioactivity & Protocols
Physicochemical Properties
Solubility Data
Calculators
Biological Data

Product Description

7α,25-dihydroxycholesterol (7α,25-OHC) is a potent and specific agonist and endogenous ligand of the orphan GPCR receptor EBI2 (GPR183). 7α, 25-dihydroxycholesterol has a strong effect on activating EBI2 (EC50=140 pM; Kd=450 pM). 7α, 25-dihydroxycholesterol works as a chemokine to guide the migration of B cells, T cells, and dendritic cells.

Biological Activity I Assay Protocols (From Reference)

ln Vitro	At a half-maximum effective concentration of roughly 500 pM, 7α,25-dihydroxycholesterol (7α,25-OHC) externally increases the migration of B and T cells in EBI2-expressing animals, but has no impact on EBI2-deficient cells [1].
ln Vivo	Restoring 7α,25-dihydroxycholesterol (1 μM; 1.5 hours) makes normal or EBI2-deficient B cells less homing to splenic follicular regions, desensitizing them [1].
References	[1]. Oxysterols direct B-cell migration through EBI2. Nature. 2011 Jul 27;475(7357):519-23. [2]. Oxysterols direct immune cell migration via EBI2. Nature. 2011 Jul 27;475(7357):524-7.
Additional Infomation	7alpha,25-dihydroxycholesterol is a 7alpha-hydroxy steroid, a 25-hydroxy steroid, an oxysterol and a 3beta-hydroxy-Delta(5)-steroid. It has a role as a human metabolite. It is functionally related to a cholesterol. 7alpha,25-Dihydroxycholesterol has been reported in Homo sapiens with data available.

These protocols are for reference only. InvivoChem does not independently validate these methods.

Physicochemical Properties

Molecular Formula	C27H46O3
Molecular Weight	418.652348995209
Exact Mass	418.344
CAS #	64907-22-8
PubChem CID	11954197
Appearance	White to off-white solid powder
Density	1.1±0.1 g/cm3
Boiling Point	548.5±35.0 °C at 760 mmHg
Melting Point	235 - 236 °C
Flash Point	230.5±20.5 °C
Vapour Pressure	0.0±3.3 mmHg at 25°C
Index of Refraction	1.550
LogP	5.75
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	5
Heavy Atom Count	30
Complexity	669
Defined Atom Stereocenter Count	9
SMILES	C[C@H](CCCC(C)(C)O)[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2[C@@H](C=C4[C@@]3(CC[C@@H](C4)O)C)O)C
InChi Key	BQMSKLCEWBSPPY-IKVTXIKFSA-N
InChi Code	InChI=1S/C27H46O3/c1-17(7-6-12-25(2,3)30)20-8-9-21-24-22(11-14-27(20,21)5)26(4)13-10-19(28)15-18(26)16-23(24)29/h16-17,19-24,28-30H,6-15H2,1-5H3/t17-,19+,20-,21+,22+,23-,24+,26+,27-/m1/s1
Chemical Name	(3S,7S,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]phenanthrene-3,7-diol
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)

Solubility Data

Solubility (In Vitro)	DMSO : ~4.55 mg/mL (~10.87 mM) Ethanol :< 1 mg/mL
Solubility (In Vivo)	Solubility in Formulation 1: 5 mg/mL (11.94 mM) in 15% Cremophor EL 85% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. 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	2.3886 mL	11.9432 mL	23.8863 mL
	5 mM	0.4777 mL	2.3886 mL	4.7773 mL
	10 mM	0.2389 mL	1.1943 mL	2.3886 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.

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In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)

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Working concentration： mg/mL；

Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.

Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O，mix and clarify.

Note： (1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.

Biological Data

Identification, structure, and pharmacological characterization of 7α, 25-OHC a: Bioactivity profile measured by mobilization of intracellular calcium of fractions from septic sheep liver extracts tested on EBI2 expressing cells (blue) or tested on a control cell line (green) expressing an unrelated GPCR called CCRL2. Activity is plotted relative to an ATP response of an endogenous ATP receptor. b: Chemical structure of 5-cholesten-3β, 7α, 25-triol (7α, 25-OHC). c: Calcium mobilization: Dose response curve induced by several related oxysterols in a CHO cell line stably expressing EBI2. Data for 7α, 25-OHC on the parental cell line lacking expression of EBI2 is given as (7α, 25-OHC, no EBI2). Data shown are from a duplicate experiment. d: Radioligand binding assay: Displacement of 10 nM 3H-7α, 25-OHC bound to membranes from EBI2 expressing CHO cells by increasing concentrations of unlabeled oxysterols. Data from quadruplicate experiments are shown.[2]. Hannedouche S, et al. Oxysterols direct immune cell migration via EBI2. Nature. 2011 Jul 27;475(7357):524-7.

Oxysterol-mediated immune cell migration a: Migration of EBV-infected B cells towards 7α, 25-OHC and closely related oxysterols. b: Migration of RS11846 cells towards 7α, 25-OHC and closely related oxysterols. c: Oxysterol-mediated RS11846 cell migration in the presence (red) or absence (blue) of pertussis toxin (PTX). d: Migration of bone marrow-derived dendritic cells (BMDC) from wild type (blue) or Ebi2(−/−) mice (red). Data for all four experiments were conducted in a transwell assay system with duplicate (a,b,c) or quadruplicate (d) samples.[2]. Hannedouche S, et al. Oxysterols direct immune cell migration via EBI2. Nature. 2011 Jul 27;475(7357):524-7.

CH25H expression regulates EBI2 bioactivity in vivo and is required for mounting a T-dependent antibody response a: Quantitative PCR analysis of Ch25h and Cyp7b1 transcript abundance in the indicated tissues or cell preparations, relative to HPRT. b: Bioassay of spleen extracts from Ch25h(−/−) or wild type animals using a reporter cell line (M12) with or without EBI2. c: Bioassay of spleen extracts from mice reconstituted with bone marrow transduced with a CH25H expressing retroviral vector (CH25H-Thy1.1) or a control vector (Thy1.1). d: Spleen sections from the indicated mice after immune challenge (day 2). Visualization of antigen-sepcific transferred (IgMa, blue) B cells in Ch25h(+/+) mice indicates that many of the activated B cells have moved to the back of the follicle and interfollicular regions, near Moma1+ marginal metallophilic macrophages (brown). In the Ch25h(−/−), the activated B cells largely fail to move to these regions and instead remain near the follicle/T cell zone interface or move into the follicle. Dashed lines indicate the follicle (F) T zone (T) boundary, identified in serial sections using IgD staining. These images are representative of multiple sections from three mice of each genotype. e: Fraction of day 2 activated B cells located in the outer follicle. Sections were stained as in d and IgMa cells within ~70um of the Moma1+ cells were enumerated and divided by the total number of IgMa cells in the follicle. Each point corresponds to an individual follicle. Enumeration was performed on sections from three mice. f: Migration of antigen-specific (MD4) B cells and non-cognate B cells from spleens of day 2 immunized wild type mice in response to 7α, 25-OHC. g: Plasma cell response in Ch25h(+/+) and Ch25h(−/−) mice at day 5 following immunization with sheep red blood cells. Left profiles show spleen cells stained for the indicated markers with plasma cells identified as B220loSyndecanhi and further stained to detect intracellular IgG1. Numbers indicate percent of total cells in the indicated gate. Right bar graph shows a summary of the data for 9 mice of each type (mean±SE). *p<0.05, **p<0.01 (unpaired student's T-test).[2]. Hannedouche S, et al. Oxysterols direct immune cell migration via EBI2. Nature. 2011 Jul 27;475(7357):524-7.