Glucosylceramide Analysis Service

Creative Proteomics offers a specialized glucosylceramide metabolomics analysis service that enables:

  • Absolute quantification of GlcCer molecular species
  • Isomer-specific resolution of GlcCer vs GalCer
  • Pathway-level profiling of glycosphingolipid metabolism
  • Cross-sample compatibility for cells, tissues, plasma, and exosomes

We support mechanistic studies, metabolic flux analysis, and lipid biomarker validation with high data precision and bioinformatics-ready outputs.

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What You Will Receive

  • Quantitative GlcCer profiles (C16–C26 chain variants)
  • Raw + normalized data tables (absolute or relative units)
  • Pathway heatmaps and metabolic summaries
  • Isomer differentiation reports (GlcCer vs GalCer)
  • Custom result interpretation + visual analytics (on request)
  • What We Provide
  • Advantages
  • Technology Platform
  • Sample Requirements
  • Demo
  • FAQs
  • Case Study

What Is Glucosylceramide?

Glucosylceramide (GlcCer) is a fundamental glycosphingolipid consisting of a ceramide backbone linked to a glucose molecule. It contributes to membrane stability, supports lipid raft assembly, and acts as a biosynthetic precursor for more complex glycosphingolipids such as lactosylceramides and gangliosides. As a core component of the sphingolipid pathway, GlcCer plays structural and regulatory roles in a variety of cellular environments.

Its dynamic profile—determined by acyl chain length, saturation, and subcellular localization—offers valuable information for studying membrane composition, lipid-mediated signaling, and glycosylation patterns.

Why Analyze Glucosylceramide?

Analyzing GlcCer helps reveal the biochemical state of glycosphingolipid metabolism and membrane-related lipid processes. Its quantification and structural characterization can support:

  • Tracking changes in sphingolipid biosynthetic or salvage pathways
  • Investigating membrane composition and lipid microdomain behavior
  • Monitoring cellular responses to environmental or experimental stimuli
  • Differentiating isomeric species such as GlcCer and GalCer with biological relevance
  • Supporting lipid flux studies and metabolic profiling in engineered systems

Glucosylceramide Analysis Service Offered by Creative Proteomics

  • Quantitative Profiling of Glucosylceramide Species: Absolute and relative quantification of GlcCer with varying acyl chain lengths (e.g., C16:0, C18:0, C20:0, C24:1) using isotopically labeled internal standards.
  • Comprehensive Mapping of Related Metabolites: Measurement of upstream and downstream metabolites including ceramide, lactosylceramide, glucosylsphingosine, and sphingosine to characterize flux through the glycosphingolipid pathway.
  • Isomer-Specific Differentiation of Hexosylceramides: Separation and identification of GlcCer vs. GalCer using MS/MS-based fragmentation patterns and retention time indexing.
  • Integrated Targeted and Untargeted Metabolomics: Simultaneous acquisition of targeted GlcCer data and broader sphingolipid profiles to support pathway enrichment and hypothesis generation.
  • Bioinformatic Pathway Mapping and Flux Analysis: Annotation and integration of GlcCer data within curated metabolic networks (e.g., KEGG, HMDB) to support lipid pathway reconstruction and quantitative flux interpretation.
  • Broad Sample Compatibility: Validated protocols for multiple sample types, including plasma, serum, cerebrospinal fluid (CSF), tissue homogenates, cell lysates, exosomes, and model organisms.
  • Multi-Omics Integration Support: Cross-platform integration with proteomic, transcriptomic, and lipidomic datasets to enable systems-level interpretation of GlcCer-related biological processes.

List of Detected Glucosylceramide and Related Metabolites

Compound NameClassDescription / Role
Glucosylceramides (GlcCer)GlycosphingolipidsPrimary targets with fatty acid chains such as:
GlcCer(d18:1/16:0)GlcCerCommon saturated species
GlcCer(d18:1/18:0)GlcCerAbundant in mammalian tissues
GlcCer(d18:1/20:0)GlcCerLonger-chain variant
GlcCer(d18:1/22:0)GlcCerTissue-enriched form
GlcCer(d18:1/24:0)GlcCerMajor storage molecule
GlcCer(d18:1/24:1)GlcCerUnsaturated form linked to metabolic stress
GlcCer(d18:0/16:0)GlcCerDihydro variant
GlcCer(d18:1/26:1)GlcCerVery long-chain unsaturated form
Glucosylsphingosine (Lyso-GlcCer)Lyso-sphingolipidDeacylated GlcCer; indicator of lysosomal storage dysregulation
Ceramides (precursors)SphingolipidsCore building blocks of GlcCer biosynthesis
Cer(d18:1/16:0) to Cer(d18:1/24:1)CeramidesSubstrates for glucosylation
Cer(d18:0/24:0), Cer(d18:0/20:0)CeramidesDihydroceramide forms
Lactosylceramides (LacCer)GlycosphingolipidsImmediate downstream product of GlcCer
LacCer(d18:1/16:0) to LacCer(d18:1/24:1)LacCerFound in membranes and signaling vesicles
Hexosylceramides (total)GlycosphingolipidsIncludes both GlcCer and GalCer isomers
GalCer(d18:1/18:0), GalCer(d18:1/24:1)GalactosylceramidesDifferentiated by MS/MS fragmentation
Sphingoid BasesBackbone lipidsMetabolites in degradation or recycling
Sphingosine (d18:1), Sphinganine (d18:0)Sphingoid basesDownstream of ceramide
Sphingomyelins (SM)PhosphosphingolipidsCompeting metabolic fates
SM(d18:1/16:0) to SM(d18:1/24:1)SphingomyelinsRelated structural lipids
UDP-glucoseNucleotide sugarDonor for glucosylation of ceramide
Other Relevant Lipid Intermediates

GlcCer-phosphateRare intermediatePossible under stress conditions
Ceramide-1-phosphate (C1P)Signaling lipidInflammatory signaling molecule
Sphingosine-1-phosphate (S1P)Signaling lipidAngiogenesis and immune signaling

Advantages of Glucosylceramide Assay

  • High Sensitivity: Detection limit as low as 0.5 ng/mL for major GlcCer species using isotopically labeled standards and optimized MRM acquisition.
  • Wide Quantification Range:Dynamic linear range from 0.5 ng/mL to 10 µg/mL, enabling accurate quantification in both trace-level and high-abundance samples.
  • Isomer-Specific Resolution:Differentiation of GlcCer vs GalCer and other hexosylceramide isomers based on MS/MS fragmentation patterns and retention time profiles.
  • Comprehensive Pathway Coverage: Simultaneous profiling of 50+ glycosphingolipid-related metabolites, including precursors (ceramides), downstream products (lactosylceramides), and degradation intermediates.
  • Cross-Matrix Compatibility: Validated for use with plasma, serum, CSF, tissues, cell lysates, and exosomes, ensuring versatility across research models.
  • Reproducibility and Technical Precision: Intra-assay CVs typically below 10%, with external QC materials applied for batch consistency.

Workflow for Glucosylceramide Analysis Service

Glucosylceramide Analysis Workflow

Technology Platform for Glucosylceramide Analysis Service

Mass Spectrometry

SCIEX QTRAP® 6500+ LC-MS/MS

Utilized for high-sensitivity targeted quantification of GlcCer species with multiple reaction monitoring (MRM) and isotope-labeled internal standards.

Thermo Scientific™ Q Exactive™ HF Orbitrap

Deployed for untargeted and semi-targeted profiling of GlcCer and related sphingolipids, offering high-resolution accurate mass detection (HRAM) and isomer differentiation.

Chromatographic Separation

Waters ACQUITY UPLC System

Equipped with BEH Amide and C18 columns for HILIC and reverse-phase separation, optimized for glycosphingolipid retention and resolution.

SCIEX Triple Quad™ 6500+

SCIEX Triple Quad™ 6500+ (Figure from Sciex)

Thermo Scientific™ Q Exactive™ HF Orbitrap

Q Exactive™ HF Orbitrap (Figure from Thermo)

Waters ACQUITY UPLC System

Waters ACQUITY UPLC System (Figure from Waters)

Sample Requirements for Glucosylceramide Analysis Service

Sample TypeMinimum Volume/WeightStorage ConditionNotes
Fruit Juice / Wine≥ 1.0 mL4°C (short-term), -20°C (long-term)Filtered; avoid preservatives if possible
Fermentation Broth≥ 0.5 mL-20°CCentrifuge to remove cells/debris; provide fermentation stage info
Plant Tissue (Fresh)≥ 100 mgFlash-frozen at -80°CIndicate species, part (e.g., leaf, fruit), and growth condition
Plant Tissue (Dry)≥ 50 mgRoom temp (sealed, dry)Grind to fine powder recommended
Plasma / Serum≥ 200 µL-80°CEDTA preferred; avoid hemolysis
Food Product (Solid)≥ 2.0 g4°C or -20°CHomogenized sample preferred; indicate composition
Standard Solution≥ 0.5 mL (optional)4°CIf customer-prepared standards are supplied

Demo Results

Bar chart showing GlcCer molecular species levels in plasma and tissue samples.

Quantitative profiling of GlcCer species across plasma and tissue samples.

MS/MS comparison of GlcCer and GalCer showing isomer-specific fragment ions.

MS/MS spectra illustrating distinct fragmentation patterns of GlcCer and GalCer.

Metabolic pathway diagram with heatmap overlay for GlcCer and related lipids.

Pathway-level heatmap of glycosphingolipid metabolism based on relative metabolite abundances.

PCA scatter plot showing sample clustering based on GlcCer-related metabolite profiles.

PCA score plot revealing metabolic separation between control and treated groups.

FAQ of Glucosylceramide Analysis Service

Can I submit different types of samples within the same project?

Yes. We accept mixed sample types (e.g., plasma, tissue, cells) within the same project. However, matrix effects and normalization strategies may differ—please indicate sample type clearly on submission.

Do I need to add internal standards before sending samples?

No. Internal standards are added during sample preparation at our facility. If you've already spiked standards, please inform us in advance to avoid overestimation or interference.

Does hemolysis or lipemia affect GlcCer quantification?

Yes. Hemolyzed or lipemic samples may introduce background noise or interfere with accurate GlcCer profiling. Please avoid hemolysis during collection and note any visible sample quality issues.

Can you work with low-abundance or precious samples?

Yes. Our platform supports high-sensitivity detection. If your sample is limited in volume or quantity, contact us—we can evaluate feasibility and adjust extraction protocols accordingly.

How should I prepare and ship tissue or cell pellet samples?

Flash-freeze immediately after collection, store at −80°C, and ship on dry ice. For cell pellets, wash with cold PBS before freezing. Include protein or wet weight data if available.

Will I receive information on related metabolites beyond GlcCer?

Yes. In addition to GlcCer species, our analysis provides data on ceramides, lactosylceramides, glucosylsphingosine, and sphingoid bases—enabling interpretation across the sphingolipid pathway.

How do you ensure separation of isobaric species like GlcCer(d18:1/18:1) vs GalCer(d18:1/18:1)?

We use retention time indexing with validated reference standards and MS/MS fragmentation patterns to distinguish GlcCer from GalCer and other isobars.

What kind of normalization or quantification approach do you use?

We apply isotope-labeled internal standards for absolute quantification when available, and normalize across sample input or protein content to ensure data comparability.

Can I integrate this lipidomics data with my proteomics or transcriptomics dataset?

Yes. We offer exportable, normalized metabolite matrices suitable for multi-omics integration. Bioinformatics support is also available if needed.

Will the result file include both normalized and raw ion intensity data?

Yes. Our reports include both raw peak areas and normalized concentrations, along with calibration curve performance and QC metrics.

Learn about other Q&A.

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Glucosylceramide Analysis Service Case Study

Title: Glucosylceramide is essential for Heartland and Dabie bandavirus glycoprotein-induced membrane fusion
Journal: PLOS Pathogens
Published: 2023

  • Background
  • Methods
  • Results
  • Reference

Heartland virus (HRTV) and Dabie bandavirus (DBV) are emerging tick-borne pathogens associated with hemorrhagic fever and high mortality. These viruses rely on Gn/Gc glycoproteins for entry into host cells, yet the precise host lipid components enabling this fusion process have remained unclear.

Sphingolipids—especially glycosphingolipids such as glucosylceramide (GlcCer)—are known to play regulatory roles in membrane structure and viral lifecycle processes. This study investigates whether GlcCer is specifically required for viral membrane fusion and entry.

The authors used genome-wide CRISPR-Cas9 screening in human 293T cells to identify host dependency factors for HRTV infection. Hits from the screen were validated through:

  • Targeted gene knockouts (UGCG, SPTLC2, UGT8, B4GALT5/6)
  • Chemical inhibition of GlcCer synthesis (e.g., NB-DNJ)
  • Quantitative lipidomics analysis using supercritical fluid chromatography–tandem mass spectrometry (SFC-MS/MS)
  • Exogenous lipid rescue experiments with GlcCer, GalCer, and LacCer
  • Membrane fusion assays and protein-lipid binding studies (SPR, BLI)

Creative Proteomics' Contribution:

The study's reliance on quantitative glycosphingolipid profiling, especially absolute GlcCer quantification and metabolic flux interpretation, demonstrates the critical role of:

  • Targeted Glucosylceramide LC-MS/MS analysis
  • Differentiation of GlcCer vs GalCer species
  • Pathway-level lipidomics of ceramide–GlcCer–LacCer axis

Creative Proteomics can support such research through:

  • High-sensitivity detection of GlcCer and related metabolites
  • Sample compatibility with cell lysates, knockout models, and viral systems
  • Quantification of metabolic changes upon gene editing or drug treatment
  • The CRISPR screen identified multiple genes in the sphingolipid biosynthesis pathway (e.g., UGCG, SPTLC2) as essential for HRTV infection.
  • Genetic or chemical inhibition of UGCG (GlcCer synthase) significantly reduced viral infectivity.
  • Supplementation with GlcCer, but not GalCer or LacCer, rescued viral entry in UGCG-deficient cells.
  • SFC-MS/MS lipidomics showed GlcCer depletion upon NB-DNJ treatment or gene knockout.
  • Biophysical assays confirmed direct binding of viral Gc protein to GlcCer, enabling membrane insertion and fusion.
  • Mutation of D841 in the Gc lipid-binding pocket abolished GlcCer interaction and viral entry.
  • Downstream glycosphingolipids (e.g., LacCer) were not required for infection; instead, GlcCer accumulation correlated with enhanced infectivity.

Bar graphs and heatmap showing reduced HRTV replication and GlcCer levels in UGCG and SPTLC2 knockout 293T cells; viral infectivity assays confirm impaired entry.Glucosylceramide (GlcCer) is essential for HRTV infection efficiency.
(A–B) Knockout of SPTLC2 or UGCG significantly reduces HRTV replication in 293T cells, as shown by qPCR. (C) Heatmap displaying intracellular GlcCer and GalCer levels across knockout lines. (D–E) Infectivity assays (TCID₅₀) show marked reduction in viral titer in UGCG- and SPTLC2-deficient cells compared to controls.

Reference

  1. Xia, Tian, et al. "Glucosylceramide is essential for Heartland and Dabie bandavirus glycoprotein-induced membrane fusion." PLoS pathogens 19.3 (2023): e1011232. https://doi.org/10.1371/journal.ppat.1011232

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Metabolomics Sample Submission Guidelines

Download our Metabolomics Sample Preparation Guide for essential instructions on proper sample collection, storage, and transport for optimal experimental results. The guide covers various sample types, including tissues, serum, urine, and cells, along with quantity requirements for untargeted and targeted metabolomics.

Metabolomics Sample Submission Guidelines
* For Research Use Only. Not for use in diagnostic procedures.
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