SPAC18B11.02c Antibody

What is SPAC18B11.02c antibody and what organism does it target?

SPAC18B11.02c antibody is a polyclonal antibody that targets the SPAC18B11.02c protein, which is found in Schizosaccharomyces pombe (strain 972 / ATCC 24843), commonly known as fission yeast . This antibody is designed specifically to recognize epitopes of this particular yeast protein and is primarily used for research applications investigating S. pombe protein expression and function.

What is the difference between polyclonal and monoclonal antibodies for SPAC18B11.02c detection?

Polyclonal SPAC18B11.02c antibodies (like the one listed in the catalog) are derived from multiple B cell lineages and recognize various epitopes on the SPAC18B11.02c protein, offering broader recognition capabilities but potentially lower specificity . Monoclonal antibodies, in contrast, are derived from a single B cell clone and recognize a single epitope, providing higher specificity but potentially lower sensitivity. For S. pombe research, polyclonal antibodies are often preferred for initial detection experiments due to their robust signal, while monoclonal antibodies might be preferred for experiments requiring higher specificity.

How is SPAC18B11.02c antibody typically validated for research applications?

SPAC18B11.02c antibodies undergo multiple validation steps similar to other research antibodies . Validation typically includes:

• Specificity testing through western blotting against S. pombe lysates

• Cross-reactivity assessment against closely related species

• Functional verification in relevant applications (ELISA, western blot)

• Independent confirmation using orthogonal methods such as mass spectrometry

• Knockout/knockdown validation when feasible using CRISPR-Cas9 or RNAi systems

What are the primary applications for SPAC18B11.02c antibody in fission yeast research?

The SPAC18B11.02c antibody has been validated for several experimental applications including:

• Western blot (WB) analysis for detecting the native protein in cell lysates

• Enzyme-linked immunosorbent assay (ELISA) for quantitative protein detection

• Potential applications in immunocytochemistry (though specific validation data may be limited)

• Protein localization studies when combined with relevant microscopy techniques

• Protein interaction studies when used in co-immunoprecipitation experiments

What are the recommended protocols for western blotting with SPAC18B11.02c antibody?

For optimal western blot results with SPAC18B11.02c antibody:

• Prepare S. pombe cell lysates using either mechanical disruption (glass beads) or enzymatic methods (zymolyase treatment)

• Use standard SDS-PAGE with 10-12% polyacrylamide gels

• Transfer proteins to PVDF or nitrocellulose membranes

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

• Incubate with primary SPAC18B11.02c antibody at optimized dilutions (typically 1:500 to 1:2000)

• Use appropriate species-specific secondary antibody (anti-rabbit HRP if using rabbit polyclonal)

• Develop using chemiluminescence and document results

How can researchers optimize ELISA protocols for SPAC18B11.02c detection?

For ELISA optimization with SPAC18B11.02c antibody :

• Coat plates with purified recombinant SPAC18B11.02c protein or S. pombe cell extracts

• Block with appropriate blocking buffer (typically 3-5% BSA)

• Prepare a dilution series of the antibody to determine optimal concentration

• Use titration experiments to determine linear range of detection

• Include appropriate positive and negative controls

• Consider sandwich ELISA approach if higher specificity is required, using two antibodies targeting different epitopes

How can researchers verify the specificity of SPAC18B11.02c antibody in their experiments?

Researchers can verify SPAC18B11.02c antibody specificity through multiple approaches :

• Parallel analysis using genetic knockouts or knockdowns of the target gene

• Independent validation using orthogonal methods (e.g., mass spectrometry)

• Testing with recombinant SPAC18B11.02c protein as a positive control

• Cross-validation using two differentially raised antibodies against the same target

• Testing against related yeast species to assess cross-reactivity

• Pre-absorption tests with purified antigen to confirm signal specificity

What controls should be included when using SPAC18B11.02c antibody?

Essential controls for experiments with SPAC18B11.02c antibody include:

• Positive control: Wild-type S. pombe extracts known to express the target protein

• Negative control: Either knockout/knockdown samples or non-related yeast species

• Loading control: Detection of a housekeeping protein (e.g., actin) to ensure equal loading

• Secondary antibody-only control: To assess background staining

• Peptide competition assay: Pre-incubation with immunizing peptide should abolish specific signal

• Isotype control: Using matched isotype immunoglobulin to assess non-specific binding

How do post-translational modifications affect SPAC18B11.02c antibody recognition?

Post-translational modifications (PTMs) can significantly impact SPAC18B11.02c antibody recognition:

• Phosphorylation, glycosylation, or other modifications may mask or alter epitopes

• Some antibodies may preferentially recognize modified or unmodified forms of the protein

• To assess PTM impact, researchers should:

  • Compare detection patterns under different physiological conditions

  • Use phosphatase or glycosidase treatments to remove specific modifications

  • Consider using modification-specific antibodies if studying particular PTMs

  • Compare results across multiple detection methods

What are common causes of weak or absent signal when using SPAC18B11.02c antibody?

When troubleshooting weak or absent signals:

• Protein expression: Verify target protein expression under your experimental conditions

• Extraction method: Ensure your extraction protocol effectively solubilizes the target protein

• Antibody concentration: Test a range of primary and secondary antibody dilutions

• Incubation conditions: Optimize temperature and duration of antibody incubations

• Detection system: Ensure chemiluminescence reagents are fresh and working properly

• Epitope accessibility: Consider native vs. denaturing conditions if epitope may be masked

• Sample degradation: Use appropriate protease inhibitors during extraction

How can researchers resolve non-specific binding issues with SPAC18B11.02c antibody?

To reduce non-specific binding:

• Optimize blocking conditions by testing different blocking agents (BSA, non-fat milk, commercial blockers)

• Increase wash stringency by adjusting salt concentration or adding detergents

• Titrate antibody concentration to find optimal signal-to-noise ratio

• Pre-absorb antibody with lysates from non-target species

• Consider alternative buffer systems for both primary and secondary antibody incubations

• Use more specific detection methods like immunoprecipitation followed by mass spectrometry

What methods can be used to quantify SPAC18B11.02c protein levels accurately?

For accurate quantification of SPAC18B11.02c protein:

• Use quantitative western blotting with:

  • Standard curves using recombinant protein

  • Digital imaging systems rather than film exposure

  • Housekeeping protein normalization

• Quantitative ELISA assays with:

  • Standard curves of known concentrations

  • Technical replicates for statistical validation

• Advanced methodologies:

  • Mass spectrometry with isotope-labeled standards

  • Automated western systems (e.g., Jess, Wes) for higher reproducibility

How can SPAC18B11.02c antibody be used in protein interaction studies?

For protein interaction studies:

• Co-immunoprecipitation (Co-IP):

  • Use SPAC18B11.02c antibody to pull down the target protein and interacting partners

  • Analyze precipitates by western blot or mass spectrometry

  • Consider crosslinking for transient interactions

• Proximity ligation assay (PLA):

  • Combine SPAC18B11.02c antibody with antibodies against potential interacting proteins

  • Visualize interactions in situ with fluorescence microscopy

• Pull-down assays:

  • Use recombinant SPAC18B11.02c protein as bait

  • Validate interactions detected with the antibody

What advanced microscopy techniques can be combined with SPAC18B11.02c antibody?

Advanced microscopy applications include:

• Super-resolution microscopy:

  • STED (Stimulated Emission Depletion)

  • STORM (Stochastic Optical Reconstruction Microscopy)

  • Requires highly specific antibodies and appropriate fluorophore conjugation

• Live cell imaging:

  • May require cell-permeable antibody fragments

  • Consider nanobody development for improved intracellular accessibility

• Correlative light and electron microscopy (CLEM):

  • Combines immunofluorescence with ultrastructural analysis

  • Requires special fixation and embedding protocols

How can researchers develop custom validation strategies for SPAC18B11.02c antibody in novel applications?

For developing custom validation strategies :

How can SPAC18B11.02c antibody be incorporated into multi-omics research approaches?

Integration strategies for multi-omics research:

• Combine immunoprecipitation with RNA-seq (RIP-seq) to identify RNA interactions

• Use ChIP-seq approaches if SPAC18B11.02c has DNA-binding properties

• Couple antibody-based purification with mass spectrometry for proteomics analysis

• Integrate data across transcriptomic, proteomic, and metabolomic platforms

• Utilize systems biology approaches to contextualize SPAC18B11.02c function within broader cellular networks

What are the considerations for using SPAC18B11.02c antibody in advanced protein engineering applications?

When using SPAC18B11.02c antibody in protein engineering:

• Epitope mapping to understand antibody binding regions

• Consideration of SpyTag/SpyCatcher systems for modular antibody construction

• Potential development of bispecific antibodies using technologies like SpyLock

• Application in protein tagging and purification systems

• Assessment of antibody fragments (Fab, scFv) for specific applications requiring smaller recognition molecules

How can researchers validate SPAC18B11.02c antibody for cross-species applications?

For cross-species validation:

• Perform sequence homology analysis across species of interest

• Test antibody against lysates from multiple species in western blot

• Use heterologous expression systems to express orthologs

• Consider epitope conservation through structural biology approaches

• Validate in knockout/knockdown systems across different species when feasible