25-Hydroxycholesterol
Based on 28 publication(s) in Google Scholar
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.
- Purity: 99.65%
- CAS No.: 2140-46-7
- Formula: C27H46O2
- Molecular Weight:402.65
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Storage:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 6 months , -20°C, 1 month
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Purity & Documentation
- References
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Publications Citing Use of MedChemExpress (MCE) 25-Hydroxycholesterol
More- Mol Cancer. 2025 May 26;24(1):151. [Abstract]
- Cell Metab. 2022 Oct 4;34(10):1548-1560.e6. [Abstract]
- Nat Aging. 2026 Feb;6(2):316-328. [Abstract]
- Mol Cell. 2023 Aug 17;83(16):3010-3026.e8. [Abstract]
- Nat Commun. 2025 Jan 31;16(1):1217. [Abstract]
- Nano Today. 2025 Oct.
- Phytomedicine. 2025 May 24:143:156904. [Abstract]
- Phytomedicine. 2024 Jun:128:155313. [Abstract]
- Cell Chem Biol. 2024 May 16;31(5):920-931.e6. [Abstract]
- J Agric Food Chem. 2025 Dec 29. [Abstract]
- Chem Biol Interact. 2024 Aug 26:111214. [Abstract]
- J Ethnopharmacol. 2022 May 23:290:115101. [Abstract]
- Life Sci. 2023 Aug 15:327:121698. [Abstract]
- Org Chem Front. 15 Feb 2022.
- Int Immunopharmacol. 2021 Jul:96:107643. [Abstract]
- J Med Virol. 2025 Jul;97(7):e70495. [Abstract]
- Microbiol Spectr. 2022 Dec 21;10(6):e0219822. [Abstract]
- Microbiol Spectr. 2021 Dec 22;9(3):e0172521. [Abstract]
- Vet Res. 2025 Jan 31;56(1):23. [Abstract]
- Viruses. 2022 Feb 8;14(2):353. [Abstract]
- Vet Res. 2021 Jun 26;52(1):95. [Abstract]
- Viruses. 2020 Jun 10;12(6):628. [Abstract]
- Arch Biochem Biophys. 2026 Jun:780:110790. [Abstract]
- Life Sci Alliance. 2022 Jan 4;5(3):e202101256. [Abstract]
- Vet Microbiol. 2024 Jan:288:109948. [Abstract]
- Biochem Biophys Res Commun. 2020 Feb 19;522(4):862-868. [Abstract]
- medRxiv. 2025 May 31:2025.05.27.25328359. [Abstract]
- bioRxiv. 2025 Jun 23:2025.06.17.660196. [Abstract]
Customer Validation & Images
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RT-PCR
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Flow Cytometry
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Cell Proliferation/Viability Assay
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IP
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WB
All Endogenous Metabolite Isoforms
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Biological Activity
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Human Endogenous Metabolite |
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Cell Line
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Type | Value | Description | References |
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| CHO-K1 | IC50 |
0.3 μM
Compound: 25-hydroxycholesterol
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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
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[PMID: 21561152] |
| HEK293 | EC50 |
2.3 μM
Compound: 11, 25-HC
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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
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[PMID: 23830695] |
| HEK293 | EC50 |
3.3 μM
Compound: 1, 25HC
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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
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[PMID: 24928400] |
| HEK293 | EC50 |
>10000 nM
Compound: 39
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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
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[PMID: 31390523] |
| HEK293 | EC50 |
>10000 nM
Compound: 39
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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
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[PMID: 31390523] |
| HEK293 | EC50 |
0.63 μM
Compound: 25HC
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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
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[PMID: 31902650] |
| HepG2 | IC50 |
0.3 μM
Compound: 25-hydroxy-cholesterol
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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
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[PMID: 8057282] |
| Vero | IC50 |
>128 μM
Compound: 7
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Cytotoxicity against african green monkey Vero cells by tetrazolium dye reduction assay
Cytotoxicity against african green monkey Vero cells by tetrazolium dye reduction assay
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[PMID: 24084159] |
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.
Chemical Information
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CAS No. 2140-46-7
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Appearance Solid
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Molecular Weight 402.65
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Formula C27H46O2
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Color White to off-white
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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
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Synonyms
25-OHC
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Structure Classification
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Initial Source
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Publications (28)
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Journal Impact Factor
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Most Recent
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Mol Cancer
GP73-dependent regulation of exosome biogenesis promotes colorectal cancer liver metastasis. [Abstract]2025 May 26;24(1):151. PMID: 40414849 -
Cell Metab
PIDDosome-SCAP crosstalk controls high-fructose-diet-dependent transition from simple steatosis to steatohepatitis. [Abstract]2022 Oct 4;34(10):1548-1560.e6. PMID: 36041455
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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Nat Aging
Comparative analysis of senolytic drugs reveals mitochondrial determinants of efficacy and resistance. [Abstract]2026 Feb;6(2):316-328. PMID: 41611832 -
Mol Cell
FAM120A couples SREBP-dependent transcription and splicing of lipogenesis enzymes downstream of mTORC1. [Abstract]2023 Aug 17;83(16):3010-3026.e8. PMID: 37595559
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.
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Nat Commun
Lymphatic-derived oxysterols promote anti-tumor immunity and response to immunotherapy in melanoma. [Abstract]2025 Jan 31;16(1):1217. PMID: 39890772
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.
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Phytomedicine
Disruption of cholesterol homeostasis triggers NLRP3-cGAS-STING axis-dependent hepatic fibrosis and honokiol intervention effects. [Abstract]2025 May 24:143:156904. PMID: 40449452 -
Phytomedicine
Alisol B regulates AMPK/mTOR/SREBPs via directly targeting VDAC1 to alleviate hyperlipidemia. [Abstract]2024 Jun:128:155313. PMID: 38520833 -
Cell Chem Biol
Transient hydroxycholesterol treatment restrains TCR signaling to promote long-term immunity. [Abstract]2024 May 16;31(5):920-931.e6. PMID: 38759618 -
J Agric Food Chem
Genistein Ameliorates Rifampicin-Undermined Hepatic Cholesterol Efflux via the CH25H-LXRα-ABCA Pathway. [Abstract]2025 Dec 29. PMID: 41459645 -
Chem Biol Interact
25-Hydroxycholesterol induces oxidative stress, leading to apoptosis and ferroptosis in extravillous trophoblasts. [Abstract]2024 Aug 26:111214. PMID: 39197811
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.
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J Ethnopharmacol
2022 May 23:290:115101. PMID: 35151834 -
Life Sci
Cordycepin reprogramming lipid metabolism to block metastasis and EMT via ERO1A/mTOR/SREBP1 axis in cholangiocarcinoma. [Abstract]2023 Aug 15:327:121698. PMID: 37080351 -
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Int Immunopharmacol
25-Hydroxycholesterol mitigates hepatic ischemia reperfusion injury via mediating mitophagy. [Abstract]2021 Jul:96:107643. PMID: 33878616 -
J Med Virol
Bergamottin, A Natural Bioactive Compound, Inhibits Dabie Bandavirus Infection In Vitro and In Vivo. [Abstract]2025 Jul;97(7):e70495. PMID: 40673712 -
Microbiol Spectr
25-Hydroxycholesterol Mediates Cholesterol Metabolism to Restrict Porcine Deltacoronavirus Infection via Suppression of Transforming Growth Factor β1. [Abstract]2022 Dec 21;10(6):e0219822. PMID: 36314946 -
Microbiol Spectr
Two Cholesterol Recognition Amino Acid Consensus Motifs of GP64 with Uncleaved Signal Peptide Are Required for Bombyx mori Nucleopolyhedrovirus Infection. [Abstract]2021 Dec 22;9(3):e0172521. PMID: 34937190 -
Vet Res
The cholesterol metabolite 25-hydroxycholesterol suppresses porcine deltacoronavirus via lipophagy inhibition and mTORC1 modulation. [Abstract]2025 Jan 31;56(1):23. PMID: 39891192 -
Viruses
Screening of Botanical Drugs against SARS-CoV-2 Entry Reveals Novel Therapeutic Agents to Treat COVID-19. [Abstract]2022 Feb 8;14(2):353. PMID: 35215943 -
Vet Res
CRISPR/Cas9-based generation of a recombinant double-reporter pseudorabies virus and its characterization in vitro and in vivo. [Abstract]2021 Jun 26;52(1):95. PMID: 34174954 -
Viruses
Broad-Spectrum Host-Based Antivirals Targeting the Interferon and Lipogenesis Pathways as Potential Treatment Options for the Pandemic Coronavirus Disease 2019 (COVID-19). [Abstract]2020 Jun 10;12(6):628. PMID: 32532085 -
Arch Biochem Biophys
Selective mitochondrial damage and dysfunction in cholesterol-exposed neuronal cells: Role of mitochondrial lipid peroxidation. [Abstract]2026 Jun:780:110790. PMID: 41825807 -
Life Sci Alliance
Necrotic debris and STING exert therapeutically relevant effects on tumor cholesterol homeostasis. [Abstract]2022 Jan 4;5(3):e202101256. PMID: 34983824 -
Vet Microbiol
The activation of liver X receptors in Madin-Darby bovine kidney cells and mice restricts infection by bovine viral diarrhea virus. [Abstract]2024 Jan:288:109948. PMID: 38113573 -
Biochem Biophys Res Commun
Combinatorial screening of a panel of FDA-approved drugs identifies several candidates with anti-Ebola activities. [Abstract]2020 Feb 19;522(4):862-868. PMID: 31806372 -
medRxiv
OSBPL11 is an African-specific locus associated with 25-hydroxyvitamin D concentrations and cardiometabolic health. [Abstract]2025 May 31:2025.05.27.25328359. PMID: 40492082 -
bioRxiv
2025 Jun 23:2025.06.17.660196. PMID: 40667025
Solvent & Solubility
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)
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.
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.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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.
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.
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
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).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL.
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
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
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Data Sheet (274 KB)
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SDS (422 KB)
- English - EN (422 KB)
- Français - FR (422 KB)
- Deutsch - DE (422 KB)
- Norwegian - NO (422 KB)
- Español - ES (422 KB)
- Swedish - SV (422 KB)
- Italian - IT (422 KB)
- Portuguese - PT (422 KB)
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Handling Instructions (2659 KB)
References
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.
| 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 |
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