ST6GALNAC1 Antibody, FITC conjugated

What is ST6GALNAC1 and why is it a valuable target for cancer research?

ST6GALNAC1 (ST6 N-acetylgalactosaminide alpha-2,6-sialyltransferase 1) is a protein sialyltransferase specifically expressed in goblet cells that plays a key role in intestinal host-commensal homeostasis . It catalyzes the formation of sialyl-Tn (S-Tn) antigen by conjugating sialic acid with an alpha-2-6 linkage to N-acetylgalactosamine (GalNAc) glycan chains linked to serine or threonine in glycoproteins .

The importance of ST6GALNAC1 in cancer research has been established through multiple studies showing that it is:

• Highly expressed in colorectal cancer stem cells/cancer initiating cells (CR-CSCs/CICs)

• Associated with enhanced sphere-forming ability and chemotherapeutic resistance

• Linked to poor prognosis when expressed in advanced-stage cancers (Stage III and IV)

• Involved in Akt pathway activation in cooperation with galectin-3

For comprehensive studies, researchers should consider analyzing ST6GALNAC1 in conjunction with cancer stem cell markers like CD44, as ST6GALNAC1 was found to sialylate CD44, potentially affecting its function in tumor progression .

What are the optimal applications for ST6GALNAC1 Antibody, FITC conjugated?

Based on manufacturer specifications and research literature, ST6GALNAC1 Antibody, FITC conjugated is primarily optimized for:

The antibody has been used successfully for detecting human ST6GALNAC1, with some cross-reactivity reported for mouse and rat samples in western blot applications . For applications requiring dual labeling, the FITC conjugation is advantageous as it eliminates the need for secondary antibodies and reduces background signal in multicolor experiments.

How should samples be prepared for immunofluorescence studies using ST6GALNAC1 Antibody, FITC conjugated?

For optimal immunofluorescence results with ST6GALNAC1 Antibody, FITC conjugated:

• Fixation:

  • For cell lines: 4% paraformaldehyde for 15-20 minutes at room temperature

  • For tissue sections: Formalin-fixed, paraffin-embedded sections with appropriate antigen retrieval

• Permeabilization:

  • 0.1-0.5% Triton X-100 or 0.5% saponin for 10 minutes

  • Note that ST6GALNAC1 is a transmembrane protein located in the Golgi apparatus, requiring adequate permeabilization

• Blocking:

  • 5-10% normal serum (from species not related to primary antibody) in PBS with 0.1% Tween-20 for 1 hour

  • Include 0.1% bovine serum albumin to reduce non-specific binding

• Antibody Dilution:

  • Start with manufacturer's recommended dilution (typically 1:100-1:500)

  • Optimization may be required based on expression levels in specific samples

• Counterstaining:

  • DAPI for nuclear visualization

  • Consider golgi markers (e.g., GM130) for co-localization studies

Researchers should note that overfixation can mask epitopes and reduce staining intensity, while insufficient permeabilization may prevent access to intracellular antigens.

What controls should be included when working with ST6GALNAC1 Antibody, FITC conjugated?

A robust experimental design with ST6GALNAC1 Antibody, FITC conjugated should include the following controls:

• Positive Control:

  • Cell lines with known ST6GALNAC1 expression (e.g., HepG2, as indicated in Western blot data)

  • Colorectal cancer tissue sections, particularly those with goblet cells

• Negative Controls:

  • Isotype control antibody (rabbit IgG-FITC) at the same concentration

  • Samples treated with siRNA targeting ST6GALNAC1 (knockdown verification)

  • Normal colon tissue (low expression comparative control)

• Technical Controls:

  • Autofluorescence control (unstained sample)

  • Single-color controls for compensation when performing multicolor flow cytometry

  • Secondary antibody-only control if using indirect detection methods

• Validation Controls:

  • Samples treated with competitive blocking using the immunogen peptide

  • Correlation with RT-PCR or Western blot data to confirm expression levels

The search results indicate that sphere-cultured colorectal cancer cells show higher ST6GALNAC1 expression than adherent-cultured cells, making these valuable comparative controls for cancer stem cell research .

How can ST6GALNAC1 Antibody, FITC conjugated be used to investigate cancer stem cells?

ST6GALNAC1 Antibody, FITC conjugated offers several methodological approaches for studying cancer stem cells:

• Flow Cytometry Protocol:

  • Harvest cells using non-enzymatic dissociation methods to preserve surface epitopes

  • Fix with 2% paraformaldehyde (10 minutes) and permeabilize with 0.1% saponin

  • Stain with ST6GALNAC1 Antibody, FITC conjugated alongside established CSC markers:

    • CD44 (APC-conjugated)

    • ALDH activity (using ALDEFLUOR assay)

  • Sort positive populations for functional assays

• Sphere Formation Assay Correlation:

  • Isolate ST6GALNAC1-high and ST6GALNAC1-low populations by FACS

  • Plate in ultra-low attachment plates in serum-free media supplemented with growth factors

  • Quantify sphere formation after 7-14 days

  • Research shows ST6GALNAC1-high populations form significantly more spheres

• Chemoresistance Analysis:

  • Treat ST6GALNAC1-positive and negative populations with chemotherapeutic agents (e.g., 5-FU)

  • Assess viability using MTT or similar assays

  • Studies have demonstrated that ST6GALNAC1 overexpression increases resistance to 5-FU

• In vivo Tumorigenicity:

  • Inject varying numbers of ST6GALNAC1-high and ST6GALNAC1-low cells into immunocompromised mice

  • Monitor tumor formation and growth

  • Calculate tumor-initiating cell frequency using limiting dilution analysis

  • Research indicates ST6GALNAC1 knockdown significantly reduces tumor formation

The integrated approach combining these methods provides comprehensive assessment of the functional relationship between ST6GALNAC1 expression and cancer stem cell properties.

How does ST6GALNAC1 contribute to the PI3K/Akt signaling pathway and how can this be studied?

ST6GALNAC1 has been shown to activate the PI3K/Akt signaling pathway in cooperation with galectin-3, contributing to cancer stem cell maintenance and chemoresistance . The following methodological approach can be used to investigate this relationship:

• Signaling Pathway Analysis:

  • Western blot analysis of key signaling molecules:

    • Phospho-Akt (Ser473)

    • Total Akt

    • Phospho-S6

    • Galectin-3

  • Compare between ST6GALNAC1-overexpressing cells, knockdown cells, and controls

  • Quantify band intensities for statistical comparison

• Galectin-3 Interaction Study:

  • Perform siRNA knockdown of galectin-3 in ST6GALNAC1-overexpressing cells

  • Assess phospho-Akt levels by Western blot

  • Research has shown galectin-3 knockdown decreases Akt phosphorylation in ST6GALNAC1-overexpressing cells

• Inhibitor Studies:

  • Treat cells with Akt inhibitors (e.g., AZD5363)

  • Assess effects on sphere formation and expression of stemness markers (ALDH1A1, SOX2)

  • Compare responses between ST6GALNAC1-high and ST6GALNAC1-low populations

• Immunofluorescence Co-localization:

  • Use ST6GALNAC1 Antibody, FITC conjugated alongside:

    • Anti-phospho-Akt antibodies (different fluorophore)

    • Anti-galectin-3 antibodies (different fluorophore)

  • Perform confocal microscopy to assess spatial relationships

  • Quantify co-localization using appropriate software

• STn Antigen Glycosylation Analysis:

  • Immunoprecipitate with anti-STn antibody

  • Perform Western blot analysis to identify carrier proteins (CD44 has been identified as a key carrier)

  • Correlate glycosylation patterns with Akt activation status

This comprehensive approach allows researchers to elucidate the mechanistic relationship between ST6GALNAC1-mediated sialylation and Akt pathway activation in cancer stem cells.

What are the methodological considerations for multiplexing ST6GALNAC1 Antibody, FITC conjugated with other markers?

When designing multiplex immunofluorescence experiments with ST6GALNAC1 Antibody, FITC conjugated:

• Fluorophore Selection:

  • FITC emits in the green spectrum (peak ~520 nm)

  • Choose complementary fluorophores with minimal spectral overlap:

    • Red spectrum: Cy3, Texas Red, Alexa Fluor 594

    • Far Red: Cy5, Alexa Fluor 647

    • Blue spectrum: DAPI, Hoechst (for nuclear counterstain)

• Recommended Marker Combinations:

  • For cancer stem cell research:

    • ST6GALNAC1-FITC + CD44-Alexa647 + ALDH1A1-Cy3

  • For sialylation studies:

    • ST6GALNAC1-FITC + Sialyl-Tn-Cy3 + Golgi marker-Alexa647

  • For signaling pathway analysis:

    • ST6GALNAC1-FITC + phospho-Akt-Cy3 + Galectin-3-Alexa647

• Sequential Staining Protocol:

  • Fix and permeabilize samples as previously described

  • Block with 10% normal serum

  • Apply ST6GALNAC1 Antibody, FITC conjugated first (1-2 hours at room temperature)

  • Wash thoroughly (3x PBS with 0.1% Tween-20)

  • Apply unconjugated primary antibodies sequentially or as a cocktail

  • Wash thoroughly

  • Apply secondary antibodies with different fluorophores

  • Wash and counterstain with DAPI

• Image Acquisition Considerations:

  • Capture single-color controls for spectral unmixing

  • Minimize bleed-through by using sequential scanning on confocal microscopes

  • FITC is susceptible to photobleaching; image this channel first or use anti-fade mounting media

• Analysis Approaches:

  • Quantify co-expression using software like ImageJ, CellProfiler, or QuPath

  • Use segmentation algorithms to define cellular compartments

  • Calculate Pearson's correlation coefficient for co-localization studies

Researchers should optimize antibody concentrations when multiplexing, as some antibodies may require higher concentrations in multiplex settings compared to single-staining protocols.

How can ST6GALNAC1 Antibody, FITC conjugated be used to study mucin integrity in intestinal research?

ST6GALNAC1 plays a crucial role in intestinal mucin sialylation, which protects intestinal mucus against bacterial proteolytic degradation . The following methodology can be employed for investigating mucin integrity:

• Tissue Section Preparation:

  • Collect fresh intestinal tissue samples

  • Process for frozen or paraffin sections (4-6 μm thickness)

  • For paraffin sections: Perform heat-induced epitope retrieval using citrate buffer (pH 6.0)

• Dual Immunofluorescence Protocol:

  • Stain with ST6GALNAC1 Antibody, FITC conjugated

  • Co-stain with mucin markers (MUC2, MUC5AC) using differently colored fluorophores

  • Counterstain nuclei with DAPI

  • Mount with anti-fade medium

• Goblet Cell Analysis:

  • Quantify the number of goblet cells (MUC2+) expressing ST6GALNAC1

  • Compare between normal and diseased tissues (e.g., ulcerative colitis)

  • Assess correlation between ST6GALNAC1 expression and mucin layer thickness

• Bacterial Penetration Assay:

  • Perform fluorescence in situ hybridization (FISH) for bacterial 16S rRNA

  • Co-stain with ST6GALNAC1 Antibody, FITC conjugated

  • Quantify bacterial penetration depth relative to ST6GALNAC1 expression levels

  • Research has shown that decreased sialylation results in less protective phenotypes

• Ex Vivo Mucus Degradation Assay:

  • Collect intestinal explants from normal and ST6GALNAC1-deficient models

  • Incubate with bacterial proteases

  • Measure mucin degradation rate

  • Correlate with ST6GALNAC1 expression levels using the FITC-conjugated antibody

These methodologies allow for comprehensive assessment of the relationship between ST6GALNAC1-mediated sialylation and mucin integrity in intestinal homeostasis.

Common Issues and Solutions:

• Weak or No Signal:

  • Cause: Insufficient antigen expression, overfixation, inadequate permeabilization

  • Solution:

    • Optimize fixation time (reduce to 10 minutes for paraformaldehyde)

    • Increase permeabilization (0.5% Triton X-100)

    • Try different antigen retrieval methods for FFPE tissues

    • Increase antibody concentration

    • Extend incubation time to overnight at 4°C

    • Use signal amplification systems

• High Background:

  • Cause: Insufficient blocking, too high antibody concentration, non-specific binding

  • Solution:

    • Increase blocking time (2 hours) and concentration (10% serum with 1% BSA)

    • Add 0.2% Tween-20 to washing buffers

    • Reduce antibody concentration

    • Pre-absorb antibody with tissue powder

    • Include detergent in antibody diluent (0.05% Triton X-100)

• Photobleaching:

  • Cause: FITC is susceptible to rapid photobleaching

  • Solution:

    • Use anti-fade mounting media containing p-phenylenediamine or ProLong Gold

    • Minimize exposure to light during processing

    • Reduce excitation light intensity during imaging

    • Consider using Alexa Fluor 488-conjugated antibodies as an alternative (more photostable)

• Non-Specific Staining:

  • Cause: Cross-reactivity, Fc receptor binding

  • Solution:

    • Include normal serum from the same species as the sample

    • Add Fc receptor blocking reagent

    • Validate specificity with knockdown controls

    • Use isotype control to determine background levels

• Inconsistent Results:

  • Cause: Batch-to-batch variation, inconsistent sample processing

  • Solution:

    • Standardize sample collection and processing protocols

    • Include positive and negative controls in each experiment

    • Document lot numbers and validate each new antibody lot

    • Consider creating a reference sample set for standardization

Creating a systematic approach to troubleshooting with appropriate controls will help researchers achieve consistent and reliable results with ST6GALNAC1 Antibody, FITC conjugated.

How can ST6GALNAC1 Antibody, FITC conjugated be used in flow cytometry for studying cancer progression?

For effective flow cytometric analysis of ST6GALNAC1 in cancer progression studies:

• Sample Preparation Protocol:

  • Harvest cells using gentle dissociation methods (e.g., Accutase rather than trypsin)

  • For tissue samples: Create single-cell suspensions using mechanical and enzymatic dissociation

  • Fix cells in 2-4% paraformaldehyde (10 minutes, room temperature)

  • Permeabilize with 0.1% saponin in PBS (15 minutes, room temperature)

  • Block with 5% normal goat serum containing 0.1% saponin

• Staining Protocol:

  • Incubate with ST6GALNAC1 Antibody, FITC conjugated (1:100 dilution, 1 hour at room temperature)

  • Wash 3× with PBS containing 0.1% saponin

  • For multiparameter analysis, include antibodies against:

    • Cancer stem cell markers (CD44, CD133, ALDH)

    • Epithelial-mesenchymal transition markers (E-cadherin, Vimentin)

    • Proliferation markers (Ki-67)

• Data Acquisition and Analysis:

  • Set up proper compensation using single-color controls

  • Acquire at least 10,000-20,000 events per sample

  • Gate strategy:

    • FSC/SSC to exclude debris

    • Single-cell gate to exclude doublets

    • Viability dye gate (if used) to exclude dead cells

    • Analyze ST6GALNAC1 expression in relevant populations

• Experimental Designs for Cancer Progression Studies:

  • Temporal Analysis:

    • Compare early vs. late-passage cancer cells

    • Monitor changes in ST6GALNAC1 expression during disease progression

  • Treatment Response:

    • Analyze ST6GALNAC1+ vs. ST6GALNAC1- cells for chemoresistance

    • Track changes in ST6GALNAC1+ population during treatment

  • Metastatic Potential:

    • Compare ST6GALNAC1 expression between primary tumors and metastatic sites

    • Research indicates ST6GALNAC1 expression correlates with metastatic potential

• Sorting Protocol for Functional Studies:

  • Sort ST6GALNAC1-high and ST6GALNAC1-low populations using a 70 μm nozzle

  • Collect in culture medium containing 20% FBS

  • Use sorted populations for:

    • Sphere formation assays

    • Invasion/migration assays

    • In vivo limiting dilution transplantation

    • RNA-seq or proteomics analysis

This comprehensive flow cytometry approach enables researchers to correlate ST6GALNAC1 expression with cancer progression and functional properties of cancer cells.

What is the relationship between ST6GALNAC1 and other sialyltransferases in cancer research?

Understanding the interplay between ST6GALNAC1 and other sialyltransferases is crucial for comprehensive glycosylation studies in cancer:

• Sialyltransferase Family Relationships:

  • ST6GALNAC1 belongs to the ST6GalNAc subfamily that transfers sialic acid to GalNAc residues via α2,6-linkage

  • Other relevant sialyltransferases in cancer research include:

    • ST3GAL1: Forms sialyl-T antigens

    • ST6GALNAC2: Prefers T and sialyl-T antigens (while ST6GALNAC1 prefers Tn antigen)

    • ST8SIA6: Forms disialic acid structures

• Differential Expression Analysis:

  • Technique: Quantitative RT-PCR panel for sialyltransferases

  • Protocol:

    • Extract RNA from normal and cancer tissues

    • Perform qRT-PCR for ST6GALNAC1, ST3GAL1, ST6GALNAC2, ST8SIA6

    • Normalize to housekeeping genes

  • Research Findings:

    • ST6GALNAC1 is often downregulated in normal tissues compared to cancer tissues

    • ST3GAL1 is typically upregulated in cancerous tissues

    • ST8SIA6 is associated with poor prognosis when highly expressed

• Functional Redundancy Assessment:

  • Approach: siRNA knockdown studies

  • Protocol:

    • Perform individual and combined knockdowns of multiple sialyltransferases

    • Assess phenotypic changes using ST6GALNAC1 Antibody, FITC conjugated and other markers

    • Measure sialyl-Tn antigen expression changes

  • Observations: ST6GALNAC1 and ST6GALNAC2 show partial functional redundancy but distinct substrate preferences

• Clinical Correlation Table:

• Multiparameter Analysis:

  • Use ST6GALNAC1 Antibody, FITC conjugated in combination with antibodies against other sialyltransferases

  • Correlate expression patterns with glycan profiles using lectin arrays

  • Integrate with patient outcome data for comprehensive biomarker assessment

Understanding these relationships enables researchers to develop more specific therapeutic strategies targeting aberrant sialylation in cancer.

How can ST6GALNAC1 Antibody, FITC conjugated be used to study the interaction between STn antigen and galectin-3?

The interaction between ST6GALNAC1-generated STn antigen and galectin-3 has significant implications for cancer progression through Akt pathway activation . Here's a methodological approach to study this interaction:

• Co-Immunofluorescence Protocol:

  • Sample Preparation:

    • Fix cells with 4% paraformaldehyde (15 minutes)

    • Permeabilize with 0.1% Triton X-100 (10 minutes)

    • Block with 5% BSA in PBS (1 hour)

  • Staining:

    • ST6GALNAC1 Antibody, FITC conjugated (1:200)

    • Anti-galectin-3 antibody with contrasting fluorophore (e.g., Cy3)

    • Anti-STn antibody with far-red fluorophore (e.g., Alexa Fluor 647)

  • Analysis:

    • Confocal microscopy with z-stack acquisition

    • Quantify triple co-localization using image analysis software

• Proximity Ligation Assay (PLA):

  • Principle: Detects proteins in close proximity (<40 nm)

  • Protocol:

    • Use unconjugated ST6GALNAC1 antibody with anti-STn and anti-galectin-3 antibodies

    • Follow manufacturer's protocol for Duolink PLA

    • Quantify interaction signals in control vs. ST6GALNAC1-overexpressing cells

• Co-Immunoprecipitation Studies:

  • Protocol:

    • Lyse cells in non-denaturing buffer

    • Immunoprecipitate with anti-STn antibody

    • Perform Western blot for galectin-3

    • Reverse immunoprecipitation with anti-galectin-3

    • Blot for STn-carrying proteins (e.g., CD44)

  • Research Finding: CD44 has been identified as a key carrier of STn antigen (~130 kDa)

• Functional Assays following Disruption of Interaction:

  • siRNA Approach:

    • Knockdown galectin-3 in ST6GALNAC1-overexpressing cells

    • Assess Akt phosphorylation status

    • Measure sphere formation capacity

    • Research shows galectin-3 knockdown decreases Akt phosphorylation and stemness

  • Inhibitor Approach:

    • Treat cells with galectin-3 inhibitors (e.g., TD139)

    • Assess effects on ST6GALNAC1-mediated phenotypes

• Glycan Engineering:

  • CRISPR-mediated knockout of ST6GALNAC1

  • Rescue experiments with:

    • Wild-type ST6GALNAC1

    • Catalytically inactive mutant

  • Analyze impact on:

    • STn antigen formation

    • Galectin-3 binding

    • Akt pathway activation

This comprehensive approach allows researchers to dissect the molecular mechanisms underlying the functional interaction between ST6GALNAC1-generated glycans and galectin-3 in cancer progression.

What experimental approaches can be used to validate the specificity of ST6GALNAC1 Antibody, FITC conjugated?

Ensuring antibody specificity is crucial for reliable research outcomes. Here are methodological approaches to validate ST6GALNAC1 Antibody, FITC conjugated:

• Genetic Validation Approaches:

  • siRNA Knockdown:

    • Transfect cells with ST6GALNAC1-specific siRNAs (at least 2 different sequences)

    • Confirm knockdown by qRT-PCR

    • Perform flow cytometry or immunofluorescence with the antibody

    • Expect significant signal reduction in knockdown cells

  • CRISPR/Cas9 Knockout:

    • Generate ST6GALNAC1 knockout cell lines

    • Validate knockout by genomic sequencing and Western blot

    • Compare antibody staining between wild-type and knockout cells

    • Complete absence of signal in knockout cells confirms specificity

• Overexpression Validation:

  • Protocol:

    • Transfect cells with ST6GALNAC1 expression vector

    • Confirm overexpression by qRT-PCR and Western blot

    • Compare antibody signal between control and overexpressing cells

    • Expect increased signal intensity in overexpressing cells

  • Controls:

    • Include empty vector transfection

    • Use cell lines with known low endogenous expression

• Peptide Competition Assay:

  • Protocol:

    • Pre-incubate antibody with excess immunizing peptide (10-100× molar excess)

    • In parallel, incubate antibody with unrelated peptide

    • Perform standard staining protocol

    • Specific signal should be blocked by the immunizing peptide but not by unrelated peptide

• Cross-Reactivity Assessment:

  • Against Related Proteins:

    • Test against cells overexpressing other ST6GALNAC family members

    • Compare staining patterns between ST6GALNAC1-6

    • Quantify signal specificity ratio

  • Against Multiple Species:

    • Test reactivity with human, mouse, and rat samples

    • Document cross-reactivity for accurate experimental planning

• Technical Validation:

  • Multiple Detection Methods:

    • Compare results from FITC-conjugated antibody with unconjugated primary + FITC-secondary approach

    • Correlation between methods supports antibody specificity

  • Antibody Titration:

    • Perform serial dilutions (1:50 to 1:1000)

    • Plot signal-to-noise ratio

    • Determine optimal concentration where specific signal is maximized and background is minimized

Implementing these validation approaches provides robust evidence for antibody specificity and strengthens the reliability of research findings using ST6GALNAC1 Antibody, FITC conjugated.

How can ST6GALNAC1 Antibody, FITC conjugated be used in patient-derived xenograft (PDX) models for cancer research?

Patient-derived xenograft (PDX) models offer valuable platforms for studying ST6GALNAC1 in cancer progression and treatment response. Here's a methodological framework:

• PDX Model Development and Characterization:

  • Tissue Processing Protocol:

    • Process fresh tumor samples within 2 hours of resection

    • Confirm ST6GALNAC1 expression in original patient tumor using immunohistochemistry

    • Implant tumor fragments subcutaneously in immunodeficient mice

    • Harvest and repassage tumors at 500-1000 mm³

  • Characterization:

    • Perform flow cytometry on dissociated PDX tumors using ST6GALNAC1 Antibody, FITC conjugated

    • Compare ST6GALNAC1 expression between patient tumor and PDX passages

    • Monitor expression stability across passages

• Tumor Heterogeneity Analysis:

  • Multiparameter Flow Cytometry:

    • Dissociate PDX tumors into single cells

    • Stain with ST6GALNAC1 Antibody, FITC conjugated and:

      • Cancer stem cell markers (CD44, CD133)

      • Differentiation markers

      • Human-specific markers (to distinguish from mouse stroma)

    • Sort ST6GALNAC1-high and ST6GALNAC1-low populations for functional assays

  • Spatial Distribution Analysis:

    • Perform multiplex immunofluorescence on frozen PDX sections

    • Map ST6GALNAC1 expression relative to hypoxic regions, vasculature, and invasive fronts

• Treatment Response Studies:

  • Protocol:

    • Establish PDX cohorts with documented ST6GALNAC1 expression profiles

    • Administer standard chemotherapy regimens

    • Harvest tumors at defined timepoints

    • Analyze changes in ST6GALNAC1 expression post-treatment

  • Analysis:

    • Correlate baseline ST6GALNAC1 expression with treatment response

    • Track emergence of ST6GALNAC1-high populations during treatment

    • Research suggests ST6GALNAC1-high cells may contribute to chemoresistance

• Targeted Therapy Approaches:

  • Combination Strategy:

    • Combine conventional chemotherapy with:

      • Akt pathway inhibitors (based on ST6GALNAC1-Akt connection)

      • Galectin-3 inhibitors

      • Glycosylation inhibitors

    • Monitor ST6GALNAC1-high population dynamics during treatment

  • Analysis Method:

    • Flow cytometry with ST6GALNAC1 Antibody, FITC conjugated

    • Immunofluorescence of tumor sections

    • qRT-PCR for expression changes

• Ex Vivo Assays with PDX-derived Cells:

  • Protocol:

    • Establish short-term cultures from PDX tumors

    • Sort ST6GALNAC1-high and ST6GALNAC1-low populations

    • Perform drug sensitivity testing, sphere formation, and invasion assays

    • Compare functional properties between populations

This comprehensive approach leverages PDX models to investigate ST6GALNAC1's role in cancer progression and treatment response, potentially identifying vulnerabilities that could be targeted therapeutically.

What are the considerations for quantitative analysis of ST6GALNAC1 expression using the FITC-conjugated antibody?

Accurate quantification of ST6GALNAC1 expression is essential for comparative studies. Here's a comprehensive methodology: