svopl Antibody

What is SVOPL and why is it studied in research?

SVOPL (SVOP-Like) is a membrane transporter protein that shares structural similarities with SVOP (synaptic vesicle 2-related protein). Research on SVOPL focuses on understanding its biological functions in cellular transport mechanisms, particularly in human tissues. The protein contains several functional domains, with antibodies typically targeting specific amino acid regions such as AA 240-269 in the middle region of the protein . Research interest in SVOPL stems from its potential role in cellular transport processes and possible implications in various physiological and pathological conditions.

What types of SVOPL antibodies are available for research applications?

Several types of SVOPL antibodies are available for research purposes, varying in the targeted epitope region and conjugation status:

• Antibodies targeting the middle region (AA 240-269)

• Antibodies targeting AA 73-129 region

• Unconjugated antibodies for flexible application

• Conjugated antibodies with various labels including:

  • HRP (Horseradish Peroxidase) for enhanced detection sensitivity

  • FITC (Fluorescein isothiocyanate) for fluorescence applications

  • Biotin for amplification strategies

  • APC (Allophycocyanin) for flow cytometry

  • PE (Phycoerythrin) for multicolor applications

What experimental validations should be performed when using a new SVOPL antibody?

When using a new SVOPL antibody, researchers should conduct several validation experiments:

• Specificity testing using positive controls (tissues/cells known to express SVOPL)

• Negative controls (tissues/cells with confirmed absence of SVOPL)

• Western blot analysis to confirm binding to a protein of expected molecular weight

• Titration experiments to determine optimal working concentration

• Cross-reactivity testing with related proteins

• Comparison with other validated SVOPL antibodies targeting different epitopes

How does epitope selection influence SVOPL antibody performance in different applications?

The epitope targeted by an SVOPL antibody significantly impacts its performance across applications. SVOPL antibodies targeting the middle region (AA 240-269) have demonstrated effectiveness in Western blotting and EIA applications . This region appears to be accessible in both native and denatured forms of the protein.

Antibodies against the AA 73-129 region show compatibility with ELISA, Western blotting, and immunofluorescence applications, suggesting this epitope maintains accessibility across multiple protein conformations . When selecting an SVOPL antibody, researchers should consider:

• Protein conformation in their experimental system (native vs. denatured)

• Potential post-translational modifications near the epitope

• Accessibility of the epitope in complex samples

• Cross-reactivity profile of antibodies targeting different regions

What are the optimal experimental conditions for using SVOPL antibodies in Western blot applications?

For optimal Western blot results with SVOPL antibodies, researchers should consider:

• Sample preparation: Complete denaturation of membrane proteins like SVOPL often requires:

  • Strong detergents (e.g., SDS)

  • Reducing agents (e.g., β-mercaptoethanol)

  • Heat treatment (95-100°C for 5-10 minutes)

• Antibody dilution: The manufacturer recommends determining the optimal working dilution experimentally , but typical starting dilutions for polyclonal antibodies are 1:500 to 1:2000

• Blocking conditions:

  • 5% non-fat dry milk or BSA in TBST

  • Block for 1 hour at room temperature

• Primary antibody incubation:

  • Overnight at 4°C or 2 hours at room temperature

  • Prepared in blocking buffer at optimized dilution

• Detection system:

  • HRP-conjugated secondary antibody for chemiluminescence

  • Fluorescently labeled secondary antibody for fluorescence imaging

How can researchers address potential cross-reactivity issues with SVOPL antibodies?

Cross-reactivity is a significant concern with polyclonal antibodies. For SVOPL antibodies:

• Perform bioinformatic analysis to identify proteins with sequence homology to the targeted epitope

• Include appropriate controls in experiments:

  • Tissues/cells with confirmed SVOPL knockout

  • Pre-adsorption controls using the immunizing peptide

  • Comparison of staining patterns with antibodies targeting different SVOPL epitopes

• Validation strategies:

  • Peptide competition assays to confirm specificity

  • Immunoprecipitation followed by mass spectrometry

  • siRNA-mediated knockdown of SVOPL to confirm signal reduction

The available SVOPL antibody has been tested for human reactivity , but potential cross-reactivity with other species or related proteins should be experimentally determined.

What buffer systems are recommended for storing and diluting SVOPL antibodies?

SVOPL antibodies are typically provided in PBS buffer with 0.09% sodium azide as a preservative . For optimal stability and performance:

• Storage conditions:

  • Store at -20°C for long-term storage

  • Aliquot to avoid repeated freeze-thaw cycles

  • For short-term storage (1-2 weeks), 4°C is acceptable

• Working dilution buffers:

  • For Western blotting: 5% BSA or non-fat dry milk in TBST

  • For ELISA: PBS with 1% BSA

  • For immunofluorescence: PBS with 1% BSA and 0.3% Triton X-100 (for permeabilization)

• Stability considerations:

  • Avoid repeated freeze-thaw cycles

  • Keep diluted antibody on ice during experiments

  • Do not add sodium azide to working solutions used with HRP-conjugated antibodies

What controls are essential when using SVOPL antibodies in immunoassays?

Proper controls are critical for interpreting results with SVOPL antibodies:

• Positive controls:

  • Cell lines or tissues known to express SVOPL

  • Recombinant SVOPL protein (if available)

• Negative controls:

  • Secondary antibody only (no primary antibody)

  • SVOPL-knockout or knockdown samples

  • Isotype control (rabbit IgG at the same concentration)

• Specificity controls:

  • Peptide competition/blocking with immunizing peptide

  • Comparison with other validated SVOPL antibodies

• Technical controls:

  • Loading controls for Western blotting

  • Counterstains for immunohistochemistry/immunofluorescence

How should researchers interpret and troubleshoot variable results with SVOPL antibodies?

Variability in results with SVOPL antibodies may stem from multiple factors:

• Sample-related issues:

  • Protein degradation (use fresh samples and protease inhibitors)

  • Insufficient protein denaturation for Western blotting

  • Epitope masking due to protein-protein interactions

  • Post-translational modifications affecting epitope recognition

• Antibody-related issues:

  • Loss of activity due to improper storage

  • Insufficient concentration

  • Lot-to-lot variability (common with polyclonal antibodies)

• Protocol optimization:

  • Adjust antibody concentration

  • Modify incubation times and temperatures

  • Test different blocking reagents

  • Optimize antigen retrieval methods for fixed tissues

• Systematic troubleshooting approach:

  • Isolate variables by changing one parameter at a time

  • Include appropriate controls in each experiment

  • Document batch information for reagents

  • Verify protein expression using complementary methods (qPCR, etc.)

How do SVOPL antibodies compare in performance across different conjugation formats?

Various conjugated formats of SVOPL antibodies offer different advantages:

Selection should be based on the specific application requirements, detection system availability, and need for multiplexing .

What are the emerging research applications for SVOPL antibodies beyond traditional immunoassays?

Beyond conventional applications, SVOPL antibodies are finding utility in:

• Proximity ligation assays (PLA) for studying:

  • Protein-protein interactions involving SVOPL

  • Spatial relationships between SVOPL and other cellular components

• ChIP-Seq applications:

  • For transcription factor variants of SVOPL

  • Investigation of potential nuclear roles

• Super-resolution microscopy:

  • Nanoscale localization of SVOPL in cellular compartments

  • Colocalization studies with other membrane transporters

• Single-cell analysis:

  • Mass cytometry (CyTOF) for high-dimensional analysis

  • Microfluidic antibody-based capture systems

• Therapeutic research:

  • Target validation in drug discovery pipelines

  • Mechanism of action studies for compounds affecting transport systems

What strategies can overcome detection sensitivity limitations with SVOPL antibodies?

When working with low SVOPL expression levels:

• Signal amplification methods:

  • Tyramide signal amplification (TSA) for immunohistochemistry

  • Poly-HRP secondary antibodies for Western blotting

  • Biotin-streptavidin amplification systems

• Sample enrichment techniques:

  • Immunoprecipitation before Western blotting

  • Subcellular fractionation to concentrate membrane fractions

  • Overexpression systems for positive controls

• Detection optimization:

  • Extended substrate incubation times

  • More sensitive substrates (e.g., femto-level chemiluminescent substrates)

  • Longer exposure times balanced against background development

  • Cooled CCD camera systems for digital imaging

• Antibody cocktails:

  • Combining multiple SVOPL antibodies targeting different epitopes

  • Using a pool of SVOPL antibodies from different host species

How can researchers optimize immunohistochemistry protocols for SVOPL detection in tissue samples?

For effective SVOPL detection in tissues:

• Fixation considerations:

  • Membrane proteins like SVOPL may require gentler fixation

  • Test both formalin-fixed and frozen sections

  • Consider light fixation (2-4% PFA for shorter times)

• Antigen retrieval options:

  • Heat-induced epitope retrieval (citrate buffer pH 6.0 or Tris-EDTA pH 9.0)

  • Enzymatic retrieval (proteinase K for membrane proteins)

  • Detergent permeabilization for improved antibody access

• Background reduction:

  • Endogenous peroxidase blocking (3% H₂O₂ in methanol)

  • Avidin/biotin blocking for biotin-based detection systems

  • Fc receptor blocking with normal serum

  • Longer blocking steps (2-3 hours) with 5-10% blocking reagent

• Signal development:

  • Polymer detection systems for increased sensitivity

  • DAB enhancement with metal ions (cobalt, nickel)

  • Extended development times with monitoring