SYT16 Antibody

What applications are SYT16 antibodies validated for?

SYT16 antibodies are primarily validated for Western Blotting (WB) and ELISA applications. Most commercial antibodies recommend dilutions of 1:500-1:1000 for WB and 1:10000 for ELISA . Some antibodies are additionally validated for immunocytochemistry and immunofluorescence (ICC-IF). When selecting an antibody, it's important to verify that it has been validated for your specific application. Western blotting validation typically shows detection of SYT16 at approximately 72kDa in human samples such as HepG2 cells .

What epitopes of SYT16 do commercially available antibodies target?

Commercial antibodies target multiple regions of the SYT16 protein:

• N-Terminal region antibodies

• Central region antibodies (AA 297-325)

• Full-length antibodies (AA 1-203)

• Mid-region antibodies (AA 218-267)

• N-terminal region antibodies (AA 51-100)

The epitope selection affects antibody performance across different applications, particularly when protein conformation or post-translational modifications may mask certain regions in native conditions. For detection of potentially modified forms of SYT16, using antibodies targeting different epitopes can provide complementary information.

What is the recommended storage protocol for SYT16 antibodies?

SYT16 antibodies are typically shipped at 4°C and should be stored at -20°C upon delivery . To maintain antibody activity:

• Aliquot upon receipt to minimize freeze-thaw cycles

• Store in temperature-stable conditions with consistent -20°C freezer temperatures

• Use sterile techniques when handling to prevent contamination

• Follow manufacturer storage buffer recommendations (typically PBS with 50% glycerol and 0.02% sodium azide)

• Record date of first use and monitor performance over time

What species reactivity do SYT16 antibodies demonstrate?

Most commercial SYT16 antibodies are reactive against human samples . Some antibodies show cross-reactivity with additional species:

• Human, dog, and pig (for certain AA 51-100 targeting antibodies)

• Human, dog, horse, rabbit, and rat (for certain internal region antibodies)

When working with non-human models, sequence homology analysis and empirical validation are essential before selecting an antibody.

How can researchers validate SYT16 antibody specificity for experimental systems?

Thorough validation is crucial for reliable results. For SYT16 antibodies, implement these validation strategies:

• Genetic validation:

  • CRISPR/Cas9 knockout or siRNA knockdown of SYT16

  • Overexpression controls with tagged SYT16 constructs

• Analytical validation:

  • Preabsorption with immunizing peptide

  • Testing multiple antibodies targeting different SYT16 epitopes

  • Western blot to confirm single band at expected molecular weight (72kDa)

  • Peptide competition assays

• Application-specific validation:

  • For ICC/IF: Include subcellular markers to confirm expected localization

  • For WB: Include positive control lysates (HepG2 has been validated )

  • For IHC: Include tissues with known expression patterns

What is the significance of SYT16 in lower-grade glioma (LGG), and how should antibodies be deployed in these studies?

SYT16 has been identified as a prognostic biomarker in LGG with important correlations to immune infiltration . Research findings demonstrate:

• Clinical correlations:

  • Increased SYT16 expression significantly correlates with tumor grade in LGG

  • Up-regulated SYT16 expression is an independent prognostic factor for good prognosis

• Immune infiltrate correlations:

  • SYT16 expression has significant negative correlations with infiltrating levels of B cells, CD4+ T cells, macrophages, neutrophils, and dendritic cells

For antibody-based studies of SYT16 in LGG:

• Use multiplexed IHC to simultaneously assess SYT16 and immune cell markers

• Consider dual analysis of protein (antibody-based) and mRNA (in situ hybridization) expression

• Include gradient samples representing different tumor grades to validate correlation

• Use standardized scoring systems for quantifying expression levels

• Correlate with patient outcome data to validate prognostic significance

What pathways are associated with high SYT16 expression and how can researchers investigate these connections?

Gene Set Enrichment Analysis (GSEA) identified several important pathways associated with high SYT16 expression in LGG :

GO terms enriched in high SYT16 expression phenotype:

• Single organism behavior

• Gated channel activity

• Cognition

• Transporter complex

• Ligand-gated channel activity

KEGG pathways differentially enriched:

• Neuroactive ligand-receptor interaction

• Calcium signaling pathway

• Long-term potentiation

• Type II diabetes mellitus

• Long-term depression

To investigate these connections:

• Perform co-immunoprecipitation studies using SYT16 antibodies to identify protein interaction partners

• Implement proximity ligation assays to confirm in situ protein interactions

• Use live-cell calcium imaging following SYT16 knockdown/overexpression to assess effects on calcium signaling

• Correlate SYT16 expression with channel activity using patch-clamp electrophysiology

• Develop multi-omics approaches correlating proteomic and transcriptomic data in SYT16-modulated systems

How can SYT16 antibodies be optimized for detection in fixed tissue samples?

When working with fixed tissue samples, especially brain tissue for LGG studies:

• Epitope retrieval optimization:

  • Test multiple antigen retrieval methods (heat-induced vs. enzymatic)

  • Optimize pH conditions (citrate buffer pH 6.0 vs. EDTA buffer pH 9.0)

  • Determine optimal retrieval time specific to SYT16 epitopes

• Signal amplification strategies:

  • Implement tyramide signal amplification for low-abundance detection

  • Use polymer-based detection systems for enhanced sensitivity

  • Consider quantum dot conjugates for multiplexed detection

• Validation approaches:

  • Use multiple antibodies targeting different SYT16 epitopes

  • Include positive control tissues with known SYT16 expression

  • Validate with orthogonal methods (RNA-seq, mass spectrometry)

How can SYT16 antibodies be employed to study its putative role in secretory vesicle trafficking?

SYT16 may be involved in trafficking and exocytosis of secretory vesicles in non-neuronal tissues and is calcium-independent . To investigate this function:

• Subcellular localization studies:

  • High-resolution confocal microscopy with SYT16 antibodies

  • Co-localization with vesicle markers (RAB proteins, VAMP family)

  • Super-resolution microscopy to resolve vesicular structures

• Functional trafficking assays:

  • Vesicle release assays following SYT16 knockdown/overexpression

  • Live-cell imaging with antibody fragments to track dynamics

  • FRAP experiments to assess mobility and turnover

• Structure-function relationships:

  • Use domain-specific antibodies to determine which regions mediate trafficking

  • Correlate with calcium independence using calcium chelators

  • Compare with calcium-dependent synaptotagmins (SYT1-SYT15)

What controls are essential when using SYT16 antibodies in cancer tissue studies?

For robust cancer tissue studies, particularly in prognostic biomarker research:

• Tissue controls:

  • Adjacent normal tissue samples

  • Gradient of tumor grades for correlation studies

  • Tissues with known high/low SYT16 expression

• Technical controls:

  • Isotype control antibodies matched to SYT16 antibody host species

  • Secondary antibody-only controls

  • Peptide competition controls

• Analytical controls:

  • Standardized positive cell lines (e.g., HepG2)

  • Quantitative standards for expression normalization

  • Blinded scoring by multiple observers

What methodological approaches can resolve contradictory findings in SYT16 expression studies?

When facing contradictory findings:

• Antibody validation comparison:

  • Compare epitope specifications of antibodies used in conflicting studies

  • Evaluate validation methods employed in each study

  • Test multiple antibodies in parallel on identical samples

• Protocol standardization:

  • Implement consistent tissue processing protocols

  • Standardize antigen retrieval methods

  • Use automated staining platforms to reduce technical variability

• Quantitative analysis approaches:

  • Employ digital pathology with standardized scoring algorithms

  • Use multi-parameter analysis accounting for heterogeneity

  • Implement machine learning approaches for pattern recognition

How can researchers investigate the negative correlation between SYT16 expression and immune infiltrates in LGG?

To explore this important finding :

• Spatial analysis techniques:

  • Multiplex immunofluorescence with SYT16 and immune cell markers

  • Spatial transcriptomics correlating SYT16 mRNA with immune signatures

  • Digital spatial profiling for high-dimensional analysis

• Functional validation approaches:

  • Co-culture experiments with SYT16-expressing cells and immune cells

  • Conditioned media experiments to test secreted factors

  • In vivo models with immune monitoring following SYT16 modulation

• Mechanistic investigations:

  • Cytokine/chemokine profiling following SYT16 knockdown/overexpression

  • Cell migration assays to assess immune cell recruitment

  • Signaling pathway analysis focused on immune regulation

What are the considerations for developing quantitative SYT16 assays for clinical applications?

For translational research toward clinical applications:

• Assay development considerations:

  • Select antibodies with high specificity and sensitivity

  • Establish quantitative standard curves using recombinant SYT16

  • Develop sandwich ELISA approaches for improved specificity

• Sample preparation optimization:

  • Standardize tissue processing for reproducible results

  • Evaluate effects of preanalytical variables (fixation time, storage)

  • Optimize extraction protocols for soluble vs. membrane-bound SYT16

• Clinical validation requirements:

  • Determine assay precision, accuracy, and reproducibility

  • Establish reference ranges in normal populations

  • Correlate with established prognostic markers and outcomes