SPAC4A8.10 Antibody

What is SPAC4A8.10 Antibody and what organism does it target?

SPAC4A8.10 Antibody (Product Code: CSB-PA519362XA01SXV) is a polyclonal antibody raised in rabbit against recombinant Schizosaccharomyces pombe (strain 972 / ATCC 24843) SPAC4A8.10 protein. This antibody specifically targets the SPAC4A8.10 protein in fission yeast with the UniProt identification number O14162. The antibody is produced through antigen affinity purification methods and is available in liquid form without conjugation .

The antibody is classified as IgG isotype and is intended strictly for research applications, not for diagnostic or therapeutic procedures. Its species reactivity is specifically limited to Schizosaccharomyces pombe (strain 972 / ATCC 24843), making it a specialized tool for fission yeast research .

What are the optimal storage conditions for SPAC4A8.10 Antibody?

For optimal antibody performance and longevity, SPAC4A8.10 Antibody should be stored at either -20°C or -80°C immediately upon receipt. Importantly, repeated freeze-thaw cycles should be strictly avoided as they can compromise antibody integrity and binding efficiency . The antibody is provided in a storage buffer containing:

• 0.03% Proclin 300 (preservative)

• 50% Glycerol

• 0.01M PBS, pH 7.4

These components help maintain antibody stability during storage. For researchers planning long-term studies, it is advisable to aliquot the antibody into smaller volumes upon initial thawing to minimize freeze-thaw cycles for subsequent experiments.

What validated applications exist for SPAC4A8.10 Antibody?

SPAC4A8.10 Antibody has been validated for specific research applications through rigorous testing. The confirmed applications include:

• Enzyme-Linked Immunosorbent Assay (ELISA)

• Western Blotting (WB)

These applications have been experimentally validated to ensure proper identification of the target antigen. When designing experiments utilizing this antibody, researchers should initially adhere to these validated applications before attempting additional techniques that would require further optimization and validation.

How should controls be incorporated when using SPAC4A8.10 Antibody?

Proper experimental controls are essential for generating reliable data with SPAC4A8.10 Antibody. A comprehensive control strategy should include:

Positive Controls:

• Wild-type S. pombe (strain 972 / ATCC 24843) lysates expressing SPAC4A8.10

• Recombinant SPAC4A8.10 protein (identical to the immunogen)

Negative Controls:

• SPAC4A8.10 knockout/deletion mutant lysates

• Non-target species lysates (e.g., S. cerevisiae) to confirm specificity

• Primary antibody omission control

• Secondary antibody-only control

When analyzing protein-protein interactions, as demonstrated in various antibody research approaches, including two-hybrid protein-protein interaction assays, researchers should systematically validate findings using multiple methodological approaches . For instance, when evaluating stress response pathways, confirmation through both co-immunoprecipitation and functional assays provides more robust evidence than either technique alone.

What optimization strategies are recommended for Western blotting with SPAC4A8.10 Antibody?

Western blotting with SPAC4A8.10 Antibody requires careful optimization to ensure specific detection of the target protein. Based on established antibody optimization protocols, the following systematic approach is recommended:

• Sample Preparation:

  • Lyse S. pombe cells under non-denaturing conditions to preserve epitope structure

  • Include protease inhibitors to prevent target degradation

  • Standardize protein quantification using BCA or Bradford assays

• Blocking Optimization:

  • Test multiple blocking agents (5% BSA, 5% non-fat milk, commercial blockers)

  • Determine optimal blocking time (1-2 hours at room temperature or overnight at 4°C)

• Antibody Dilution Series:

  • Perform an initial dilution series (1:500, 1:1000, 1:2000, 1:5000)

  • Evaluate signal-to-noise ratio to determine optimal concentration

• Incubation Parameters:

  • Test both room temperature (1-2 hours) and 4°C (overnight) incubation

  • Compare continuous gentle agitation versus static incubation

Similar to optimization approaches used for other antibodies, researchers should document all parameters systematically to identify conditions yielding maximum specificity with minimal background .

How can cross-reactivity concerns be addressed with SPAC4A8.10 Antibody?

Cross-reactivity assessment is crucial for antibody specificity validation, particularly with polyclonal antibodies like SPAC4A8.10. Based on established antibody validation principles, researchers should implement:

• Comparative Analysis:

  • Test reactivity against related proteins with structural homology

  • Include closely related species (e.g., other Schizosaccharomyces species)

  • Analyze potential cross-reactivity with human or other model organism proteins if conducting comparative studies

• Specificity Verification Methods:

  • Pre-absorption tests with recombinant SPAC4A8.10 protein

  • Peptide competition assays

  • Parallel testing in knockout/knockdown models

Cross-reactivity assessment approaches have been well-documented in antibody research literature, noting that comprehensive validation significantly improves experimental reliability across applications .

How can SPAC4A8.10 Antibody be incorporated into protein-protein interaction studies?

SPAC4A8.10 Antibody can serve as a valuable tool for investigating protein-protein interactions using multiple complementary techniques. Drawing from established protein interaction methodologies, researchers could implement:

• Co-immunoprecipitation (Co-IP):

  • Use SPAC4A8.10 Antibody to immunoprecipitate the target protein along with binding partners

  • Analyze precipitated complexes via mass spectrometry for unbiased interactome mapping

  • Confirm specific interactions with targeted Western blotting

• Proximity Ligation Assays:

  • Combine SPAC4A8.10 Antibody with antibodies against suspected interaction partners

  • Visualize protein-protein interactions in situ with subcellular resolution

• Two-Hybrid Validation:

  • Utilize two-hybrid systems adapted for S. pombe as described in research literature

  • Clone SPAC4A8.10 into appropriate vectors (similar to pGBKT7 or pGADT7 AD vectors)

  • Validate interactions identified through initial screening

Similar approaches have been successfully employed in studying protein interactions in stress response pathways, where antibodies were used to purify protein complexes for subsequent mass spectrometry analysis, resulting in the identification of numerous interacting partners .

What methodological considerations exist for using SPAC4A8.10 Antibody in stress response research?

Stress response research represents a significant application area for SPAC4A8.10 Antibody, particularly if the target protein functions in cellular stress pathways. Based on established stress response research protocols, researchers should consider:

• Stress Condition Optimization:

  • Test multiple stress types (temperature, oxidative, osmotic, nutritional)

  • Establish precise time-course parameters for acute versus chronic stress

  • Document stress intensity gradients for dose-response relationships

• Subcellular Localization Analysis:

  • Monitor potential translocation of SPAC4A8.10 under stress conditions

  • Combine antibody-based detection with GFP-tagged constructs for live-cell imaging

  • Implement subcellular fractionation to quantify compartment-specific distribution

• Protein Complex Dynamics:

  • Analyze stress-induced changes in SPAC4A8.10 interaction partners

  • Evaluate post-translational modifications arising during stress response

  • Compare interaction profiles between normal and stress conditions

This approach parallels methodologies used in glucocorticoid receptor stress response research, where researchers identified temperature-specific protein interactions through comparative immunoprecipitation followed by mass spectrometry analysis .

How can SPAC4A8.10 Antibody be integrated with other research techniques for comprehensive analysis?

Integrating SPAC4A8.10 Antibody with complementary techniques creates powerful research workflows for comprehensive protein characterization. Researchers can implement multi-technique approaches including:

• Combined Omics Pipeline:

  • Use antibody-based purification followed by mass spectrometry

  • Correlate protein interaction data with transcriptomics profiles

  • Integrate with metabolomics to establish functional relationships

• Structure-Function Analysis:

  • Combine antibody-detected expression patterns with mutation studies

  • Correlate protein abundance with functional assays

  • Map antibody epitopes to functional domains for structure-activity relationships

• Temporal Dynamics Analysis:

  • Synchronize cell populations to study cell-cycle-dependent changes

  • Implement time-course experiments with precisely timed sampling

  • Correlate protein levels with cellular phenotypes across developmental stages

Similar integrated approaches have been successfully employed in stress response research, where antibody-based techniques were combined with functional thermotolerance assays and gene expression analysis to establish comprehensive pathway understanding .

How should researchers address weak or absent signals when using SPAC4A8.10 Antibody?

When confronting weak or absent signals with SPAC4A8.10 Antibody, a systematic troubleshooting approach is essential. Based on established antibody troubleshooting protocols, researchers should consider:

• Protein Expression Verification:

  • Confirm target protein expression using alternative detection methods

  • Verify sample integrity through detection of housekeeping proteins

  • Consider potential post-translational modifications affecting epitope accessibility

• Technical Optimization:

  • Reduce stringency of washing steps

  • Increase antibody concentration

  • Extend incubation periods

  • Test alternative detection systems with higher sensitivity

• Sample Preparation Refinement:

  • Optimize protein extraction methods

  • Test different lysis buffers

  • Evaluate need for denaturation vs. native conditions

  • Consider epitope masking in protein complexes

This methodical approach mirrors troubleshooting strategies employed for antibodies with challenging targets, where systematic optimization successfully resolved detection issues .

What strategies can address non-specific binding with SPAC4A8.10 Antibody?

Non-specific binding represents a common challenge with polyclonal antibodies. When encountering this issue with SPAC4A8.10 Antibody, researchers should implement a targeted optimization strategy:

• Blocking Enhancement:

  • Test alternative blocking agents (casein, fish gelatin, commercial formulations)

  • Extend blocking duration

  • Add blocking agents to antibody dilution buffer

• Wash Protocol Optimization:

  • Increase wash buffer stringency (higher detergent concentration)

  • Extend washing duration and number of wash cycles

  • Consider alternative detergents (Tween-20, Triton X-100, NP-40)

• Antibody Conditioning:

  • Pre-absorb antibody with non-target proteins

  • Increase antibody dilution

  • Implement subtractive approaches using knockout/knockdown samples

These approaches align with strategies used to optimize antibody specificity in challenging systems, where methodical refinement successfully minimized background while maintaining specific signal detection .

How can researchers interpret contradictory results from different applications using SPAC4A8.10 Antibody?

Contradictory results across different applications require careful analysis and reconciliation. When confronting such discrepancies with SPAC4A8.10 Antibody, researchers should consider:

• Application-Specific Epitope Accessibility:

  • Recognize that different applications expose different protein conformations

  • Native versus denatured conditions affect epitope accessibility

  • Fixation methods can alter protein structure and antibody recognition

• Comprehensive Validation Strategy:

  • Confirm results with alternative antibodies targeting different epitopes

  • Implement orthogonal detection methods (mass spectrometry, PCR)

  • Utilize tagged protein constructs as parallel validation

• Environmental Variable Analysis:

  • Document differences in sample preparation between applications

  • Evaluate buffer composition variations

  • Consider protein complex dynamics in different experimental contexts

This analytical approach has proven effective in resolving contradictory findings in antibody-based research, where technique-specific factors were systematically identified and addressed .

What emerging technologies could enhance SPAC4A8.10 Antibody applications?

SPAC4A8.10 Antibody applications can be extended through integration with emerging technologies, significantly expanding research capabilities. Based on current trends in antibody-based research, promising directions include:

• Advanced Imaging Technologies:

  • Super-resolution microscopy for precise subcellular localization

  • Expansion microscopy for improved spatial resolution

  • Label-free detection methods for live-cell dynamics

• Single-Cell Applications:

  • Antibody-based techniques adapted for single-cell proteomics

  • Spatial transcriptomics combined with protein detection

  • Microfluidic platforms for high-throughput single-cell analysis

• Structural Analysis Integration:

  • Combining antibody epitope mapping with cryo-EM structural analysis

  • In-cell NMR with antibody validation

  • Proximity labeling approaches for interaction domain mapping

These technological integrations parallel innovative approaches being implemented across antibody research fields, where multi-modal analysis provides unprecedented insights into protein function and dynamics .

How can SPAC4A8.10 Antibody contribute to stress response pathway mapping?

SPAC4A8.10 Antibody offers significant potential for mapping stress response pathways in S. pombe, particularly when implemented within comprehensive research frameworks. Based on established stress response research methodologies, researchers could:

• Develop Interactome Maps:

  • Use antibody-based purification under various stress conditions

  • Identify condition-specific interaction partners via mass spectrometry

  • Create dynamic interaction networks across stress response timepoints

• Characterize Functional Domains:

  • Combine antibody detection with domain mutation analysis

  • Map regions essential for stress-induced interactions

  • Identify regulatory motifs through correlation of structural features with stress response

• Establish Evolutionary Conservation:

  • Compare SPAC4A8.10 interactions with homologous proteins across species

  • Identify conserved versus species-specific stress response mechanisms

  • Evaluate potential therapeutic targets in pathogenic fungi

This approach builds upon methodologies successfully employed in stress response research, where systematic protein interaction analysis revealed novel pathway components and regulatory mechanisms .

What research frontiers remain unexplored for SPAC4A8.10 Antibody applications?

Despite its validated applications, significant research opportunities remain for SPAC4A8.10 Antibody in advancing understanding of S. pombe biology. Future research directions with substantial potential impact include:

• Systems Biology Integration:

  • Multi-omics investigations correlating protein dynamics with global cellular responses

  • Network analysis identifying central regulatory hubs in stress response

  • Modeling approaches predicting phenotypic outcomes from protein interaction maps

• Comparative Model Organism Studies:

  • Cross-species investigations of conserved protein functionality

  • Evolutionary analysis of stress response pathways

  • Translation of findings to higher eukaryotic systems

• Methodological Innovations:

  • Development of SPAC4A8.10-specific biosensors

  • Implementation of optogenetic approaches for dynamic analysis

  • Integration with CRISPR-based interrogation systems

These research frontiers reflect broader trends in antibody-based biological investigation, where integrative approaches continue to reveal previously unrecognized aspects of protein function and regulation .