Cholesterol-13C2 |
Catalog No.GC64477
One-Click Copy Product Info
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콜레스테롤-13C2는 콜레스테롤로 표시된 13C입니다.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 78887-48-6
Sample solution is provided at 25 µL, 10mM.
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Cholesterol-13C2 is a 13C-labeled cholesterol. Cholesterol-13C2 serves as an internal standard in mass spectrometry and NMR analyses for accurate quantification and metabolic studies of cholesterol and related metabolites[1-4].
References:[1] Ostlund RE Jr, Matthews DE. [13C]cholesterol as a tracer for studies of cholesterol metabolism in humans. J Lipid Res. 1993 Oct;34(10):1825-31.
[2] Lehmann WD, Kessler M. Accurate quantification of nonesterified and total cholesterol in plasma and urine from crude lipid extracts by stable isotope dilution and field desorption mass spectrometry. J Clin Chem Clin Biochem. 1982 Dec;20(12):893-8.
[3] Shackleton CH, Reid S. Diagnosis of recessive X-linked ichthyosis: quantitative HPLC/mass spectrometric analysis of plasma for cholesterol sulfate. Clin Chem. 1989 Sep;35(9):1906-10.
[4] Ferezou J, Sulpice JC, Coste T, et al. Origins of neutral sterols in human feces studied by stable isotope labeling (D and 13C). Effect of phytosterols and calcium. Digestion. 1982;25(3):164-72.
Cholesterol-13C2 is used in cholesterol uptake experiments[1].
This experimental protocol is compiled based on the research content. The protocol is for reference only. Please adjust the protocol according to actual needs.
(1) Reagent preparation: Dissolve Cholesterol-13C2 in chloroform to prepare a stock solution. Use 20μL olive oil to prepare a solution containing 540μmol Cholesterol-13C2. Dissolve Cholesterol-13C2 in chloroform to prepare an internal standard solution at 0.4μg/20μL. Prepare 150mol/L sodium chloride solution. Prepare chloroform/methanol (2:1; v/v) extraction solution. Prepare hexane/isopropanol (95:5; v/v) as liquid chromatography mobile phase.
(2) Cell culture and spiking: Seed and culture Caco-2 cells to 12-14 days post-confluency monolayers.
(3) In vitro digestion and sample preparation: Take 2.0mL Ensure, 2.0mL orange juice (with or without phytosterols), and 20μL olive oil containing 540μmol Cholesterol-13C2. Perform simulated gastric and small intestinal digestion. After digestion, homogenize and centrifuge at 150000g, 4℃ for 35min. Filter the supernatant through 0.22μm to collect. Dilute with Dulbecco/s minimal essential medium at 1:4. Add to Caco-2 cell monolayers and culture for 4h (37℃, 5% CO2).
(4) Cell washing and collection: Remove medium. Wash cell monolayers twice with ice-cold phosphate-buffered saline (PBS) containing 2g/L albumin. Wash twice with ice-cold PBS without albumin. Collect cells in 1.5mL ice-cold PBS. Centrifuge at 1800rpm, 4℃ for 5min. Store cell pellets at -80℃ under nitrogen.
(5) Cholesterol extraction: Thaw cell pellets. Add 0.3mL 150mol/L sodium chloride. Sonicate on ice (setting 3, 10s). Add 1.2mL chloroform/methanol (2:1) to extract cholesterol. Vortex. Centrifuge at 2000g to accelerate phase separation. Repeat extraction twice. Combine chloroform layers (lower layers). Dry under nitrogen stream. Dissolve residue in 1mL hexane. Filter through 0.2μm membrane. Use for LC/APCI-MS/MS analysis.
Precautions:
(1) The method detection limit is 2.2pmol. The lower limit of quantification is 7.2pmol. The linear range is 7.2-1127pmol. The regression coefficient R>0.997.
(2) Cholesterol-13C2 is stable in hexane at room temperature for 2 weeks.
(3) For your safety and health, wear a lab coat and disposable gloves during operation.
References:[1] Tian Q, Failla ML, Bohn T, et al. High-performance liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry determination of cholesterol uptake by Caco-2 cells. Rapid Commun Mass Spectrom. 2006;20(20):3056-60.
| Cas No. | 78887-48-6 | SDF | |
| Formula | C2513C2H46O | M.Wt | 388.64 |
| Solubility | Ethanol : 5.56 mg/mL (14.31 mM; Need ultrasonic) | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.5731 mL | 12.8654 mL | 25.7308 mL |
| 5 mM | 514.6 μL | 2.5731 mL | 5.1462 mL |
| 10 mM | 257.3 μL | 1.2865 mL | 2.5731 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)