C2 Ceramide
Based on 2 publication(s) in Google Scholar
C2 Ceramide (Ceramide 2) is the main lipid of the stratum corneum and a protein phosphatase 1 (PP1) activator. C2 Ceramide activates PP2A and ceramide-activated protein phosphatase (CAPP). C2 Ceramide induces cells differentiation, autophagy and apoptosis, inhibits mitochondrial respiratory chain complex III. C2 Ceramide is also a skin conditioning agent that protects the epidermal barrier from water loss.
For research use only. We do not sell to patients.
- Purity : 99.85%
- CAS No.: 3102-57-6
- Formula: C20H39NO3
- Molecular Weight:341.53
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Storage:
-20°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
To place orders, for customer services and technical support, please contact: MedChemExpress USA
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Protocols
- Purity & Documentation
- References
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Help & FAQs
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Publications Citing Use of MedChemExpress (MCE) C2 Ceramide
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Customer Validation & Images
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WB
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RT-PCR
Biological Activity
Description
IC50 & Target
Protein phosphatase 1[2]
PP2A[4]
Ceramide-activated protein phosphatase (CAPP)[4]
Apoptosis[1]
Mitochondrial respiratory chain complex III[1]
Cellular Effect
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Cell Line
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Type | Value | Description | References |
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| CCRF-CEM | IC50 |
31.6 μM
Compound: C2-ceramide
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In vitro growth inhibitory effect in human T-cell acute leukemia cells CCRF-CEM
In vitro growth inhibitory effect in human T-cell acute leukemia cells CCRF-CEM
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[PMID: 11689086] |
| FL5.12 | IC50 |
33 μM
Compound: 9
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Cytotoxicity against mouse FL5.12A cells after 48 hrs by DAPI staining-based flow cytometric analysis
Cytotoxicity against mouse FL5.12A cells after 48 hrs by DAPI staining-based flow cytometric analysis
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[PMID: 27475534] |
| HCT-116 | EC50 |
24.5 μM
Compound: 3, C2-ceramide
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Cytotoxicity against HCT116 cells assessed as cell survival by SRB assay
Cytotoxicity against HCT116 cells assessed as cell survival by SRB assay
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[PMID: 17561396] |
| HepG2 | IC50 |
58 μM
Compound: 1a
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Antiproliferative activity against human HepG2 cells after 24 hrs by Alamar blue assay
Antiproliferative activity against human HepG2 cells after 24 hrs by Alamar blue assay
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[PMID: 25172146] |
| HL-60 | IC50 |
26 μM
Compound: 1a
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Antiproliferative activity against human HL60 cells after 24 hrs by Alamar blue assay
Antiproliferative activity against human HL60 cells after 24 hrs by Alamar blue assay
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[PMID: 25172146] |
| HT-29 | IC50 |
25.9 μM
Compound: C2-ceramide
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Antiproliferative activity against human HT-29 cells measured after 72 hrs by sulforhodamine B assay
Antiproliferative activity against human HT-29 cells measured after 72 hrs by sulforhodamine B assay
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[PMID: 33592537] |
| K562 | EC50 |
35.1 μM
Compound: 3, C2-ceramide
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Cytotoxicity against K562 cells assessed as cell survival by MTT assay
Cytotoxicity against K562 cells assessed as cell survival by MTT assay
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[PMID: 17561396] |
| MCF7 | IC50 |
45.41 μM
Compound: 1, C2 ceramide
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Cytotoxicity against human MCF7 cells after 24 hrs by CellTiter-Blue assay
Cytotoxicity against human MCF7 cells after 24 hrs by CellTiter-Blue assay
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[PMID: 19171486] |
| MDA-MB-231 | IC50 |
35.37 μM
Compound: 1, C2 ceramide
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Cytotoxicity against human MDA-MB-231 cells after 24 hrs by CellTiter-Blue assay
Cytotoxicity against human MDA-MB-231 cells after 24 hrs by CellTiter-Blue assay
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[PMID: 19171486] |
| NCI-H358 | EC50 |
19.6 μM
Compound: 3, C2-ceramide
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Cytotoxicity against NCI-H358 cells assessed as cell survival by SRB assay
Cytotoxicity against NCI-H358 cells assessed as cell survival by SRB assay
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[PMID: 17561396] |
| NHDF | IC50 |
66.5 μM
Compound: 1, C2 ceramide
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Cytotoxicity against NHDF after 24 hrs by CellTiter-Blue assay
Cytotoxicity against NHDF after 24 hrs by CellTiter-Blue assay
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[PMID: 19171486] |
| PC-12 | IC50 |
98 μM
Compound: 1a
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Antiproliferative activity against rat PC12 cells after 24 hrs by Alamar blue assay
Antiproliferative activity against rat PC12 cells after 24 hrs by Alamar blue assay
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[PMID: 25172146] |
| SK-BR-3 | IC50 |
49.2 μM
Compound: 1, C2 ceramide
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Cytotoxicity against human SKBR3 cells after 24 hrs by CellTiter-Blue assay
Cytotoxicity against human SKBR3 cells after 24 hrs by CellTiter-Blue assay
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[PMID: 19171486] |
In Vitro
C2 Ceramide (5 nM-200 μM; 24 hours; primary mouse osteoblasts) treatment (≤500 nM) promots osteoblast viability, whilst concentrations ≥2 μM significantly reduces osteoblast viability in a dose- and time-dependent manner[1].
C2 Ceramide increases cytoplasmic histone-associated DNA fragments by 5.7- and 11.2-fold at 50 μM and 100 μM C2 Ceramide concentrations respectively in osteoblasts. At these higher concentrations, C2 Ceramide is a potent inducer of apoptosis in osteoblasts[1].
C2 Ceramide up-regulates mRNA expression of angiogenic genes in human dental pulp cells (HDPCs) and increases the migration and capillary tube formation of endothelial cells, whereas PP1 small interfering RNA shows opposite effects. Human dental pulp cells (HDPCs) increases levels of bone morphogenetic protein 2, phosphorylation of Smad 1/5/8, and mRNA expression of runt-related transcription factor 2 and osterix[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:Primary mouse osteoblasts
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Concentration:5 nM-200 µM
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Incubation Time:24 hours
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Result:Murine osteoblasts demonstrated a dose-dependent increase in their survival rate when exposed to low concentrations of 5-500 n M. Increasing concentrations of 20-200µM caused a dose-dependent decrease in mitochondrial succinate dehydrogenase activity and osteoblast survival.
In Vivo
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Chemical Information
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CAS No. 3102-57-6
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Appearance Solid
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Molecular Weight 341.53
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Formula C20H39NO3
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Color White to off-white
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SMILES
O[C@H](/C=C/CCCCCCCCCCCCC)[C@H](CO)NC(C)=O
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Synonyms
Ceramide 2
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
-20°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
Publications (2)
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Journal Impact Factor
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Most Recent
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Neuron
Stem cell-derived neurons reflect features of protein networks, neuropathology, and cognitive outcome of their aged human donors. [Abstract]2021 Nov 3;109(21):3402-3420.e9. PMID: 34473944 -
Food Funct
A casein-derived peptide exerted immunomodulatory effects by targeting protein phosphatase 1-catalytic subunit alpha: a potential alternative to combat infections. [Abstract]2025 Aug 11;16(16):6422-6433. PMID: 40654180
C2 Ceramide purchased from MedChemExpress. Usage Cited in: Food Funct. 2025 Aug 11;16(16):6422-6433. [Abstract]
Western blot analysis for phosphorylation of proteins in NF-κB and MAPK signaling pathways in THP-1 cells treated with BCCY-1 in the presence and absence of C2 ceramide (20 μM, 1-3 h).
C2 Ceramide purchased from MedChemExpress. Usage Cited in: Food Funct. 2025 Aug 11;16(16):6422-6433. [Abstract]
The mRNA expressions of MCP-1 in BCCY-1-treated THP-1 cells incubated with or without C2 ceramide (20 μM) measured by qRT-PCR.
Solvent & Solubility
In Vitro:
Ethanol : 100 mg/mL (292.80 mM; Need ultrasonic)
DMSO : 50 mg/mL (146.40 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 (sealed storage, away from moisture). 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 (sealed storage, away from moisture). 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)
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.
Add each solvent one by one: 10% EtOH 90% Corn Oil
Solubility: 5 mg/mL (14.64 mM); Clear solution; Need ultrasonic
This protocol yields a clear solution of 5 mg/mL. 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 EtOH stock solution (50.0 mg/mL) to 900 μL Corn oil, and mix evenly.
In Vivo Dissolution Calculator
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. * In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
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.
Protocols
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Apoptosis
Apoptosis, also called programmed cell death, is generally characterized by distinct morphological characteristics.
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TUNEL staining for apoptotic DNA fragmentation
TUNEL staining detects DNA strand breaks by using terminal deoxynucleotidyl transferase to add labeled nucleotides to exposed 3′-OH DNA termini, generating either microscopic staining in fixed cells or tissue sections, or fluorescence/cytometric signal in cell suspensions. TUNEL positivity reflects DNA fragmentation but should not be interpreted alone as definitive apoptosis, because TUNEL can also label necrotic, autolytic, mechanically damaged, or DNA-repair-associated DNA breaks.
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Annexin V plus membrane-impermeant dye apoptosis staining
Annexin V-based apoptosis assays rely on the detection of phosphatidylserine (PS) externalization from the inner leaflet of the plasma membrane to the outer leaflet, an early biochemical hallmark of apoptosis. Fluorescently labeled Annexin V binds PS in a calcium-dependent manner, enabling identification of early apoptotic cells by flow cytometry or fluorescence microscopy. When combined with a membrane-impermeant DNA-binding dye (e. g. , propidium iodide), this approach allows discrimination between viable (Annexin V−/dye−), early apoptotic (Annexin V+/dye−), and late apoptotic or necrotic (Annexin V+/dye+) cell populations by assessing membrane integrity and PS exposure.
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Autophagy
Autophagy is a process in which eukaryotic cells use lysosomes to degrade their own cytoplasmic proteins and damaged organelles under the regulation of autophagy related gene (Atg). Microtubule-associated proteins light chain 3 (LC3) is recognized as autophagy marker, which transfers from cytoplasmic LC3 (LC3-I) to membrane type (LC3-II). LC3-II/I ratio could be detected by Western Blot and fluorescence microscopy.
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Apoptosis Solutions
Apoptosis is a regulated, generally non-lytic cell-death pathway that removes unwanted, damaged, infected, or abnormal cells through coordinated morphological changes, caspase activation, DNA fragmentation, and membrane remodeling. The intrinsic apoptosis pathway is controlled mainly by mitochondrial outer membrane permeabilization, BCL-2 family proteins, cytochrome c release, apoptosome formation, caspase-9 activation, and downstream executioner caspase-3/7 activation. The extrinsic apoptosis pathway is initiated by death receptors such as Fas, TNFR, and TRAIL receptors, which recruit adaptor proteins and activate caspase-8 before engaging executioner caspases or mitochondrial amplification through BID cleavage. Apoptosis is linked to many phenotypes, including cancer cell killing, tissue homeostasis, immune regulation, neurodegeneration, infection response, and treatment-induced cytotoxicity; unresolved questions include how apoptosis interacts with necroptosis, pyroptosis, ferroptos
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Lysosome and acidic-vesicle live-cell staining
Lysosome and acidic-vesicle live-cell staining detects acidic intracellular compartments by using membrane-permeant acidotropic probes that accumulate in low-pH vesicles, including lysosomes, late endosomes, autolysosomes, and acidic phagosomes. LysoTracker staining is commonly used as an intensity-based readout of acidic lysosomal compartment abundance or enlargement, while acridine orange produces green fluorescence in less concentrated compartments and red fluorescence after concentration-dependent accumulation in acidic vesicular organelles. Loss or reduction of acridine-orange red signal can be used as a readout of lysosomal membrane permeabilization or reduced acidic-vesicle integrity. This protocol is designed for live cultured cells and can be adapted for fluorescence microscopy, high-content imaging, plate-reader readout, or flow cytometry when the selected literature supports the readout. Because these dyes report acidotropic accumulation rather than lysosome identity alone,
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Macroautophagy Solutions
Macroautophagy is a conserved lysosome-dependent degradation pathway in which cytoplasmic material is sequestered into double-membrane autophagosomes and delivered to lysosomes for degradation and recycling. The pathway supports cellular homeostasis during nutrient limitation, organelle stress, protein-aggregate accumulation, infection, differentiation, and tissue remodeling by coupling cargo sequestration, autophagosome maturation, lysosomal fusion, and degradation of cargo-derived macromolecules. The core molecular sequence includes initiation by nutrient- and stress-regulated autophagy machinery, autophagosome nucleation, LC3/ATG8-family conjugation to autophagosomal membranes, cargo selection through receptors such as SQSTM1/p62, autophagosome-lysosome fusion, and lysosomal degradation. LC3 was identified as a mammalian homolog of yeast Atg8 that localizes to autophagosomal membranes after processing, and p62/SQSTM1 was shown to connect ubiquitinated cargo with autophagic degradati
Purity & Documentation
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Data Sheet (273 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Handling Instructions (2659 KB)
References
[1]. P A Hill, et al. Ceramide-induced Cell Death/Survival in Murine Osteoblasts. J Endocrinol. 2010 Aug;206(2):225-33. [Content Brief]
[2]. Ji-Youn Kim, et al. Role of Protein Phosphatase 1 in Angiogenesis and Odontoblastic Differentiation of Human Dental Pulp Cells. J Endod. 2017 Mar;43(3):417-424. [Content Brief]
[3]. R T Dobrowsky, et al. Ceramide Stimulates a Cytosolic Protein Phosphatase. J Biol Chem. 1992 Mar 15;267(8):5048-51. [Content Brief]
[4]. R T Dobrowsky, et al. Ceramide Activates Heterotrimeric Protein Phosphatase 2A. J Biol Chem. 1993 Jul 25;268(21):15523-30. [Content Brief]
[6]. Wenyuan Zhu, et al. C2-ceramide induces cell death and protective autophagy in head and neck squamous cell carcinoma cells. Int J Mol Sci. 2014 Feb 21;15(2):3336-55. [Content Brief]
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 (sealed storage, away from moisture). 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 | 2.9280 mL | 14.6400 mL | 29.2800 mL | 73.2000 mL |
| 5 mM | 0.5856 mL | 2.9280 mL | 5.8560 mL | 14.6400 mL | |
| 10 mM | 0.2928 mL | 1.4640 mL | 2.9280 mL | 7.3200 mL | |
| 15 mM | 0.1952 mL | 0.9760 mL | 1.9520 mL | 4.8800 mL | |
| 20 mM | 0.1464 mL | 0.7320 mL | 1.4640 mL | 3.6600 mL | |
| 25 mM | 0.1171 mL | 0.5856 mL | 1.1712 mL | 2.9280 mL | |
| 30 mM | 0.0976 mL | 0.4880 mL | 0.9760 mL | 2.4400 mL | |
| 40 mM | 0.0732 mL | 0.3660 mL | 0.7320 mL | 1.8300 mL | |
| 50 mM | 0.0586 mL | 0.2928 mL | 0.5856 mL | 1.4640 mL | |
| 60 mM | 0.0488 mL | 0.2440 mL | 0.4880 mL | 1.2200 mL | |
| 80 mM | 0.0366 mL | 0.1830 mL | 0.3660 mL | 0.9150 mL | |
| 100 mM | 0.0293 mL | 0.1464 mL | 0.2928 mL | 0.7320 mL |
C2 Ceramide Related Classifications
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