SPBC3B8.06 Antibody

What is SPBC3B8.06 and what is its significance in fission yeast?

SPBC3B8.06 is a protein-coding gene found in Schizosaccharomyces pombe (fission yeast), identified by the UniProt number O59714. It belongs to the comprehensive set of proteins that have been systematically characterized through phenomics approaches in recent studies of fission yeast. While still being actively researched, understanding SPBC3B8.06 contributes to our broader knowledge of conserved yeast proteins, many of which have orthologs in humans . The significance of SPBC3B8.06 aligns with the broader goal in molecular biology of characterizing poorly understood proteins even in well-studied organisms like S. pombe, where phenomics approaches have provided functional cues for thousands of genes, including many previously uncharacterized proteins .

What are the specifications of commercially available SPBC3B8.06 antibodies?

Commercial SPBC3B8.06 antibodies typically include polyclonal antibodies raised in rabbits against recombinant Schizosaccharomyces pombe (strain 972/ATCC 24843) SPBC3B8.06 protein . The antibody package generally contains three key components:

• Recombinant immunogen protein/peptide (200μg) - used as a positive control

• Pre-immune serum (1ml)

• Rabbit polyclonal antibody purified by Protein A/G

These antibodies are typically unconjugated and have been validated for applications including ELISA and Western Blot analysis . Given that they are raised against specific fission yeast proteins, they demonstrate species reactivity with yeast but may not cross-react with proteins from other organisms.

What are the recommended storage and handling conditions?

For optimal performance and longevity, SPBC3B8.06 antibodies should be stored at either -20°C or -80°C . This is consistent with standard practices for antibody storage to prevent protein degradation and maintain functional activity. When working with the antibody:

• Avoid repeated freeze-thaw cycles that can degrade antibody quality

• Aliquot the antibody into single-use volumes before freezing if multiple experiments are planned

• Allow the antibody to thaw completely at 4°C before use

• Following reconstitution (if applicable), store working dilutions at 4°C for short-term use (1-2 weeks)

• Before each use, centrifuge the antibody vial briefly to collect the solution at the bottom

These precautions help preserve antibody specificity and reactivity, particularly important for research applications where quantitative results are required.

What experimental approaches are optimized for SPBC3B8.06 antibody use?

The SPBC3B8.06 antibody has been validated primarily for ELISA and Western Blot applications , making these the preferred methodologies for experimental use. For Western Blot applications:

• Sample preparation: Extract proteins from fission yeast cultures using standard protocols with protease inhibitors

• SDS-PAGE: Separate proteins by molecular weight using 10-12% acrylamide gels

• Transfer: Transfer proteins to PVDF or nitrocellulose membranes using standard techniques

• Blocking: Block membranes with 5% non-fat milk or BSA in TBST for 1 hour at room temperature

• Primary antibody: Incubate with SPBC3B8.06 antibody (recommended dilution range: 1:500-1:2000) overnight at 4°C

• Secondary antibody: Use anti-rabbit HRP-conjugated secondary antibody for detection

• Detection: Visualize using chemiluminescence or fluorescence-based systems

For ELISA applications, the antibody can be used as a detection antibody in indirect ELISA protocols, with optimization required for different experimental setups.

How can SPBC3B8.06 antibody contribute to phenotypic profiling studies?

SPBC3B8.06 antibody can be a valuable tool in phenotypic profiling studies, particularly when integrated with the extensive phenomics data now available for fission yeast . Recent research has generated quantitative phenotype data for 3492 non-essential genes across 131 diverse conditions, with phenotypes detected for 99.6% of mutants studied . To leverage SPBC3B8.06 antibody in phenotypic studies:

• Correlation with deletion phenotypes: Use Western blot to measure SPBC3B8.06 protein levels in various genetic backgrounds and correlate with phenotypic data from deletion studies

• Localization studies: Employ immunofluorescence using SPBC3B8.06 antibody to determine protein localization patterns during different growth conditions

• Protein interaction studies: Use co-immunoprecipitation with SPBC3B8.06 antibody to identify interaction partners under conditions where specific phenotypes are observed

• Stress response studies: Monitor SPBC3B8.06 protein levels and modifications in response to the same stressors used in phenomics screens (oxidative, osmotic, heavy-metal stresses)

This approach allows researchers to connect protein-level observations with the rich phenotypic data available, potentially revealing mechanisms underlying observed phenotypes.

What control experiments should be performed to validate SPBC3B8.06 antibody specificity?

Rigorous validation of antibody specificity is crucial for reliable experimental outcomes. For SPBC3B8.06 antibody, the following validation experiments are recommended:

• Positive control: Use the provided recombinant immunogen protein (200μg) as a positive control in Western blot or ELISA to confirm antibody reactivity

• Pre-immune serum control: Compare results with the provided pre-immune serum (1ml) to identify any non-specific binding

• Knockout/knockdown validation: Test the antibody against samples from SPBC3B8.06 deletion mutants (which should show no signal) compared to wild-type controls

• Peptide competition assay: Pre-incubate the antibody with excess immunizing peptide to block specific binding sites before application to samples

• Cross-reactivity assessment: Test the antibody against proteins from related yeast species to evaluate specificity

Document these validation experiments thoroughly, as they provide critical evidence for antibody specificity that should be included in publications.

How should researchers troubleshoot inconsistent SPBC3B8.06 antibody results?

When facing inconsistent results with SPBC3B8.06 antibody, a systematic troubleshooting approach should be implemented:

Document all optimization steps systematically to establish reliable protocols for future experiments.

How can SPBC3B8.06 antibody data be integrated with machine learning predictions?

The integration of antibody-based experimental data with computational predictions represents a powerful approach in modern molecular biology research. For SPBC3B8.06:

• Validation of computational predictions: Use SPBC3B8.06 antibody to experimentally verify protein expression levels or modifications predicted by machine learning approaches such as the NET-FF predictor, which combines protein-network and protein-family data to predict Gene Ontology (GO) terms

• Training data generation: Employ quantitative Western blot data from SPBC3B8.06 antibody experiments across various conditions to generate training datasets for machine learning algorithms

• Multi-omics integration: Combine SPBC3B8.06 antibody-based proteomic data with transcriptomics and phenomics data to create integrated models of protein function prediction

• Network analysis validation: Use co-immunoprecipitation with SPBC3B8.06 antibody to validate protein-protein interactions predicted by network analysis algorithms

This integration is particularly valuable given recent advances in predicting gene functions using machine learning, which has generated 56,594 high-scoring GO predictions for fission yeast proteins, including many previously uncharacterized proteins .

What approaches enable correlation of SPBC3B8.06 function with other fission yeast proteins?

To establish functional relationships between SPBC3B8.06 and other fission yeast proteins, several complementary approaches can be employed:

• Phenotype correlation networks: Analyze the similarity of phenotypic profiles between SPBC3B8.06 deletion mutants and other gene deletions across 131 different conditions to identify functionally related proteins through "guilt by association"

• Co-immunoprecipitation studies: Use SPBC3B8.06 antibody to pull down protein complexes, followed by mass spectrometry to identify interaction partners

• Genetic interaction mapping: Combine SPBC3B8.06 deletion with other gene deletions to identify synthetic lethal or suppressor relationships, which can be visualized using protein expression studies with the antibody

• Subcellular co-localization: Perform dual-labeling immunofluorescence with SPBC3B8.06 antibody and antibodies against other proteins of interest to identify spatial relationships

• Comparative expression analysis: Measure SPBC3B8.06 protein levels across various genetic backgrounds and stress conditions, correlating with expression patterns of other proteins

These approaches collectively provide a multi-dimensional view of SPBC3B8.06's functional relationships within the cellular network.

What are the current limitations in interpreting SPBC3B8.06 functional data?

Despite advances in characterizing fission yeast proteins, several limitations remain when interpreting SPBC3B8.06 functional data:

• Incomplete functional characterization: While phenomics approaches have provided broad cues for many proteins, the specific molecular functions of SPBC3B8.06 may still not be fully characterized

• Antibody epitope considerations: The polyclonal nature of available antibodies means they recognize multiple epitopes, potentially masking or overrepresenting certain protein conformations or modified forms

• Context-dependent functions: SPBC3B8.06 may perform different functions under different conditions, making unified functional assignment challenging

• Redundancy considerations: Potential functional redundancy with other proteins may obscure phenotypes in single-gene deletion studies

• Post-translational modification detection: Current antibodies may not distinguish between modified forms of SPBC3B8.06, limiting insights into regulatory mechanisms

Addressing these limitations requires combining multiple complementary approaches and developing more specific reagents for studying this protein.

How might SPBC3B8.06 research contribute to understanding conserved cellular processes?

Fission yeast serves as an important model organism with many conserved cellular processes relevant to human biology. Research on SPBC3B8.06 could contribute to broader understanding in several ways:

• Evolutionary conservation: If SPBC3B8.06 has human orthologs, understanding its function in yeast could provide insights into conserved processes across eukaryotes

• Stress response mechanisms: Given the extensive phenotyping of fission yeast mutants under stress conditions, SPBC3B8.06 research might reveal conserved stress response pathways

• Protein interaction networks: Mapping SPBC3B8.06 interactions could reveal conserved protein complexes or pathways

• Cellular aging processes: Recent validation studies based on GO predictions have revealed new proteins involved in cellular aging in fission yeast, and SPBC3B8.06 might similarly contribute to this important biological process

• Disease-relevant processes: If SPBC3B8.06 functions in conserved processes disrupted in human diseases, this research could eventually inform therapeutic approaches

This research exemplifies how detailed characterization of seemingly obscure proteins in model organisms can contribute to fundamental understanding of cellular processes across species.