SPAC637.06 Antibody

What is SPAC637.06 and why is it studied in research?

SPAC637.06 is a protein found in Schizosaccharomyces pombe (fission yeast) with UniProt accession number P78817. While its precise biological function remains uncharacterized in public databases as of 2025, it is studied as part of broader research into fission yeast as a model organism. Fission yeast proteins are frequently investigated as models for eukaryotic cellular processes, including cell cycle regulation and DNA repair mechanisms that may be conserved across species. The SPAC637.06 protein represents one of many partially characterized components in the S. pombe proteome that requires further investigation to complete our understanding of eukaryotic cell biology.

What applications is the SPAC637.06 Antibody validated for?

The SPAC637.06 Antibody (Product Code: CSB-PA302908XA01SXV) has been validated for ELISA and Western Blot (WB) applications. For ELISA applications, the antibody has been specifically confirmed for qualitative detection of SPAC637.06 in Schizosaccharomyces pombe lysates. In Western Blot applications, the antibody has been verified to identify the antigen in fission yeast samples, though exact molecular weight data are not provided in the current documentation. The antibody has not been explicitly validated for other common applications such as immunoprecipitation, immunofluorescence, or flow cytometry, which would require additional optimization and validation by researchers.

What are the optimal storage conditions for SPAC637.06 Antibody?

The optimal storage conditions for SPAC637.06 Antibody are at either -20°C or -80°C. It is critically important to avoid repeated freeze-thaw cycles as these can significantly degrade the antibody and reduce its effectiveness in experimental applications. The antibody is supplied in a storage buffer containing 50% glycerol, 0.01M PBS (pH 7.4), and 0.03% Proclin 300 preservative, which helps maintain stability during storage. For working solutions, researchers should prepare only the amount needed for immediate use and store any remaining stock solution according to the manufacturer's recommendations to maintain antibody integrity and performance consistency across experiments.

What is the species reactivity of the SPAC637.06 Antibody?

The species reactivity of the SPAC637.06 Antibody is specifically limited to Schizosaccharomyces pombe (fission yeast). The antibody was raised against a recombinant form of the SPAC637.06 protein from Schizosaccharomyces pombe strain 972, ensuring high specificity for its target in this organism. This strict species specificity means the antibody is not expected to cross-react with proteins from other species, including related yeasts like Saccharomyces cerevisiae or mammalian systems. Researchers working with other model organisms would need to identify homologs of SPAC637.06 and obtain or develop separate antibodies specific to those proteins.

What is the composition of the storage buffer for SPAC637.06 Antibody?

The storage buffer for SPAC637.06 Antibody consists of 50% glycerol, 0.01M PBS at pH 7.4, and 0.03% Proclin 300 preservative. Each component serves a specific purpose in maintaining antibody stability: the glycerol acts as a cryoprotectant to prevent freezing damage during storage at -20°C or -80°C; the PBS maintains appropriate pH conditions critical for antibody stability; and the Proclin 300 prevents microbial contamination during storage and handling. This buffer formulation is optimized to preserve antibody function while allowing for convenient aliquoting and storage. Researchers should avoid diluting the stock solution unless preparing working dilutions for immediate use in experiments.

How can I optimize Western Blot protocols for SPAC637.06 detection in fission yeast samples?

Optimizing Western Blot protocols for SPAC637.06 detection requires systematic consideration of multiple experimental variables. For sample preparation, use fresh fission yeast cultures and prepare lysates using a buffer containing protease inhibitors to prevent protein degradation. Mechanical disruption methods like glass bead lysis are particularly effective for S. pombe due to its robust cell wall. For protein loading, determine the optimal amount through titration experiments, typically starting with 20-40 μg of total protein per lane.

For blocking, use 5% non-fat dry milk or BSA in TBST (Tris-buffered saline with 0.1% Tween-20) for 1 hour at room temperature. Regarding antibody dilution, begin with manufacturer-recommended dilutions (typically 1:500 to 1:2000) and optimize as needed. For primary antibody incubation, overnight at 4°C often yields the best results with reduced background. Include thorough washing steps (3-5 washes with TBST, 5-10 minutes each) to reduce non-specific binding.

For detection, choose an appropriate secondary antibody (anti-rabbit IgG) conjugated to HRP, and use a sensitive detection reagent suitable for your imaging system. Always include both positive controls (known S. pombe samples expressing SPAC637.06) and negative controls (S. pombe knockout strains if available) to validate specificity and optimize signal-to-noise ratio.

What are the experimental considerations when using SPAC637.06 Antibody for localization studies?

When conducting localization studies with SPAC637.06 Antibody, several critical experimental considerations must be addressed for reliable results. For fixation method, in S. pombe, a formaldehyde-based fixation (3.7% formaldehyde for 30 minutes) typically works well, though optimization may be required for specific experimental needs. Permeabilization is especially important as S. pombe has a robust cell wall requiring effective permeabilization through enzymatic digestion with zymolyase or lysing enzymes followed by detergent treatment (0.1% Triton X-100).

Blocking conditions should use 5% BSA or normal serum from the species of the secondary antibody to reduce non-specific binding. Antibody concentration should be titrated to find the optimal concentration that provides specific staining with minimal background. Controls must include appropriate negative controls such as cells not expressing SPAC637.06 (if available) and secondary-only controls to distinguish true signal from background.

Consider using known organelle markers to help determine the subcellular localization of SPAC637.06 through co-localization studies. For imaging, use confocal microscopy with appropriate filter sets and imaging parameters to detect the fluorescent signal accurately. Finally, develop consistent quantification methods to analyze the distribution pattern of SPAC637.06 across different experimental conditions or genetic backgrounds.

How can I validate the specificity of SPAC637.06 Antibody in my experimental system?

Validating the specificity of SPAC637.06 Antibody requires a multi-faceted approach combining several complementary methods. Begin with Western Blot analysis to confirm the antibody detects a single band of the expected molecular weight in wild-type S. pombe lysates. If available, use SPAC637.06 knockout or knockdown strains as negative controls—the absence or reduction of signal would strongly confirm antibody specificity.

Compare signal intensity between wild-type cells and those overexpressing SPAC637.06, as an increased signal in overexpressing cells supports antibody specificity. Consider performing a peptide competition assay by pre-incubating the antibody with excess purified SPAC637.06 protein or the immunizing peptide, which should abolish or significantly reduce the signal if the antibody is truly specific.

More advanced validation can include immunoprecipitation followed by mass spectrometry to confirm the antibody pulls down SPAC637.06 protein. If possible, test the antibody against closely related proteins to assess potential cross-reactivity. Finally, ensure results are reproducible across different batches of the antibody and experimental conditions to establish robust validation of antibody specificity in your specific experimental system.

What are the recommended positive and negative controls when using SPAC637.06 Antibody?

When using SPAC637.06 Antibody, implementing appropriate controls is essential for experimental rigor and result interpretation. For positive controls, include wild-type S. pombe lysates known to express SPAC637.06; recombinant SPAC637.06 protein (similar to the immunogen used to raise the antibody); and if available, S. pombe strains with tagged or overexpressed SPAC637.06. These positive controls establish the expected signal pattern and intensity.

For negative controls, include SPAC637.06 knockout strains if available; RNA interference-mediated knockdown of SPAC637.06; lysates from organisms not expressing SPAC637.06 or homologs; secondary antibody-only controls (omitting primary antibody); and pre-immune serum controls for polyclonal antibodies. Additionally, peptide competition assays where excess antigen is used to block antibody binding can serve as specificity controls.

Technical controls should include loading controls (e.g., actin, tubulin) for Western blot normalization; sample processing controls to ensure consistent protein extraction across samples; and antibody titration series to determine optimal working concentrations for your specific application. This comprehensive control strategy ensures reliable interpretation of experimental results and validation of antibody specificity.

How can SPAC637.06 Antibody be used to investigate protein-protein interactions?

SPAC637.06 Antibody can be employed in multiple methodologies to investigate protein-protein interactions. Co-immunoprecipitation (Co-IP) represents a primary approach, where the antibody is used to pull down SPAC637.06 from cell lysates, followed by analysis of co-precipitated proteins by mass spectrometry or Western blot. Consider crosslinking before lysis to capture transient interactions that might otherwise be lost during sample processing.

For spatial analysis of interactions, proximity ligation assay (PLA) combines SPAC637.06 Antibody with antibodies against suspected interaction partners, where PLA signal indicates close proximity (< 40 nm) between proteins and provides spatial information about interactions within cells. Similarly, immunofluorescence co-localization uses SPAC637.06 Antibody alongside antibodies against potential interactors, with quantification of co-localization using appropriate statistical measures.

To validate interactions identified through these approaches, consider complementary methods such as pull-down assays with tagged proteins, using SPAC637.06 Antibody to validate pull-downs and compare interactors identified with tagged versus endogenous protein. Bimolecular fluorescence complementation (BiFC) and FRET analysis can provide additional confirmation and information about the cellular context of identified interactions, with SPAC637.06 Antibody serving as a validation tool for these results.

What are the potential cross-reactivity issues with SPAC637.06 Antibody?

Several potential cross-reactivity issues should be considered when working with SPAC637.06 Antibody. The antibody might recognize homologous proteins with similar epitopes in S. pombe or other species, especially if the epitope is located in a conserved domain. This is particularly relevant when studying protein families or when attempting to use the antibody in related yeast species.

The antibody was raised against a recombinant form of SPAC637.06, which may affect its ability to recognize the native protein versus denatured forms in different applications. This could result in differential performance between applications like Western blot (denatured protein) versus immunoprecipitation (native protein). If SPAC637.06 undergoes post-translational modifications that alter the epitope recognized by the antibody, detection may be inconsistent across different cellular states or conditions.

Additionally, high concentrations of the antibody might lead to non-specific binding to other proteins, especially in complex samples. To address these issues, always include appropriate controls, validate the antibody in your specific experimental system, consider using multiple antibodies targeting different epitopes of SPAC637.06, optimize antibody concentration to minimize non-specific binding, and potentially pre-absorb the antibody with lysates from organisms not expressing SPAC637.06 to reduce background.

How can SPAC637.06 Antibody be used in combination with other techniques to elucidate the protein's function?

SPAC637.06 Antibody can be combined with diverse techniques to comprehensively investigate the protein's function. For proteomics approaches, use immunoprecipitation followed by mass spectrometry to identify interaction partners; if SPAC637.06 has nuclear localization, consider ChIP-seq to identify potential DNA binding sites; and employ phospho-proteomics to identify changes in phosphorylation states upon SPAC637.06 perturbation.

In genetic approaches, correlate protein expression/localization (detected by the antibody) with phenotypes in genetic screens and validate genetic interactions using the antibody to confirm protein expression/depletion. For cell biology techniques, combine live-cell imaging of fluorescently tagged proteins with immunofluorescence using the antibody for validation, and use subcellular fractionation with Western blotting to determine precise localization patterns.

Time-course experiments can track SPAC637.06 expression and localization during cell cycle progression or monitor changes in response to various stresses or environmental conditions. For comparative studies, examine SPAC637.06 across different yeast strains or related species and correlate protein conservation with functional conservation. This multi-technique approach leverages the antibody as part of a broader experimental strategy to elucidate the biological role of this uncharacterized protein.

What is known about the structural domains of SPAC637.06 and how does this affect antibody binding?

Based on available information, detailed structural domain data for SPAC637.06 is limited. The protein has UniProt accession number P78817, but its precise biological function and domain organization remain uncharacterized in public databases as of 2025. Without specific structural information, several general considerations regarding antibody binding can be applied to experimental design.

The SPAC637.06 Antibody was raised against a recombinant form of the entire protein, suggesting it likely recognizes epitopes that are accessible in the denatured protein. The effectiveness of the antibody in different applications (ELISA versus Western Blot) indicates that the epitope(s) recognized are accessible in both denatured (Western Blot) and partially native (ELISA) conformations. If SPAC637.06 undergoes significant conformational changes during its functional cycle, the antibody might recognize certain states preferentially.

To better understand the relationship between SPAC637.06 structure and antibody binding, researchers should consider conducting epitope mapping to identify the specific region recognized by the antibody; analyzing the protein sequence using bioinformatics tools to predict domains; comparing with structurally characterized homologs from other organisms; and evaluating how sample preparation methods might affect protein conformation and epitope accessibility in different experimental contexts.

What is the recommended dilution range for SPAC637.06 Antibody in Western Blot applications?

While specific recommended dilution ranges for SPAC637.06 Antibody in Western Blot applications may vary between laboratories, general principles for polyclonal antibodies against yeast proteins suggest starting with a dilution range of 1:500 to 1:2000 in primary antibody buffer (typically TBST with 1-5% BSA or non-fat dry milk). Optimization through systematic titration is essential for achieving optimal results in your specific experimental system.

The optimization approach should involve performing a titration experiment using 3-4 different dilutions (e.g., 1:500, 1:1000, 1:2000, 1:5000) and evaluating signal strength and background levels at each dilution. Choose the dilution that provides the strongest specific signal with minimal background. Sample-dependent adjustments may be necessary—higher protein loading may require more dilute antibody to prevent oversaturation, while samples with low SPAC637.06 expression may require more concentrated antibody.

For incubation conditions, overnight incubation at 4°C often provides optimal results with reduced background, though room temperature incubation (1-2 hours) may work with less dilute antibody concentrations. For secondary antibody, use anti-rabbit IgG secondary antibody at manufacturer's recommended dilution (typically 1:2000 to 1:10000) and ensure it is compatible with your detection system. Document optimal conditions once established for reproducibility across experiments.

How can I troubleshoot weak or absent signals when using SPAC637.06 Antibody?

Troubleshooting weak or absent signals with SPAC637.06 Antibody requires a systematic approach addressing multiple potential issues. For sample preparation problems, ensure you're using fresh samples with protease inhibitors and avoid repeated freeze-thaw cycles; verify sufficient protein loading by increasing amount and confirming protein concentration measurement; optimize lysis methods specifically for S. pombe (e.g., glass bead disruption, enzymatic cell wall digestion); and confirm complete denaturation of samples for SDS-PAGE.

For antibody-related issues, check antibody storage conditions and avoid repeated freeze-thaw cycles; try more concentrated primary antibody dilution; consider epitope accessibility by testing different sample preparation methods; and extend primary antibody incubation time (e.g., overnight at 4°C). Technical issues might include transfer problems (ensure efficient protein transfer to membrane; consider using transfer verification stains); blocking issues (try different blocking agents; optimize blocking time); detection sensitivity (use more sensitive detection reagents; extend exposure time); and secondary antibody concerns (confirm correct species reactivity; try more concentrated dilution).

Experimental design considerations should include a known positive control expressing SPAC637.06; assessment of whether SPAC637.06 is expressed at detectable levels under your experimental conditions; confirmation that you're examining the correct molecular weight region; and consideration of whether post-translational modifications might alter antibody recognition or protein migration.

What sample preparation methods are optimal for SPAC637.06 detection?

Optimal sample preparation methods for SPAC637.06 detection in S. pombe vary depending on the application. For Western Blot analysis, collect exponentially growing cultures (OD600 = 0.5-1.0) and harvest cells by centrifugation (3,000 × g for 5 minutes), followed by washing the cell pellet with cold PBS or water. For cell lysis, mechanical disruption using glass bead lysis in an appropriate buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM EDTA, 10% glycerol) with protease inhibitor cocktail is effective for S. pombe. Consider adding 1% Triton X-100 or NP-40 for membrane protein solubilization if needed.

After lysis, clarify samples by centrifugation (14,000 × g for 10-15 minutes at 4°C) and collect the supernatant containing soluble proteins. For membrane proteins, specific extraction buffers may be required. Quantify proteins using Bradford, BCA, or similar assays for consistent loading across samples (20-40 μg per lane typical). For sample denaturation, add Laemmli sample buffer (final concentration 1×) and heat at 95°C for 5 minutes, though for membrane proteins, consider alternative temperatures (37°C or 65°C).

For immunoprecipitation applications, use milder lysis conditions with gentler detergents (0.5% NP-40 or 0.5% Triton X-100) and include protease and phosphatase inhibitors. Consider crosslinking for transient interactions (1% formaldehyde, 10 minutes) and pre-clear lysates with Protein A/G beads before adding antibody to reduce non-specific binding.

Can SPAC637.06 Antibody be used for immunoprecipitation studies?

While the available information does not explicitly mention validation of SPAC637.06 Antibody for immunoprecipitation (IP) applications, researchers can test and optimize this application using a methodological approach. For initial feasibility testing, recognize that many polyclonal antibodies that work well in Western Blot can also function in IP. Perform a small-scale test IP using standard protocols and analyze the precipitated material by Western Blot using the same antibody.

Protocol optimization should test different lysis buffers (varying salt concentration, detergent type/concentration); antibody amounts (typically 1-5 μg antibody per 0.5-1 mg protein lysate); incubation times (overnight at 4°C usually gives best results); and bead types (Protein A, Protein G, or Protein A/G beads, noting that Protein A works well for rabbit IgG).

Essential controls include normal rabbit IgG to assess non-specific binding; input sample to confirm presence of target protein in starting material; and supernatant sample to assess IP efficiency. For validation, use Western Blot to probe IP material with SPAC637.06 Antibody; consider mass spectrometry to confirm SPAC637.06 enrichment; and if interaction partners are identified, confirm interactions by reciprocal IP with antibodies against those partners.

Troubleshooting considerations include epitope accessibility (the epitope may be masked in native conformation); antibody affinity (higher affinity is generally required for IP than Western Blot); and the potential need for cross-linking to capture transient interactions.

What are the best fixation methods for immunofluorescence studies using SPAC637.06 Antibody?

While immunofluorescence applications are not specifically mentioned for SPAC637.06 Antibody in the available information, researchers can optimize fixation methods for S. pombe immunofluorescence through systematic testing. Formaldehyde fixation (3.7-4% formaldehyde in PBS for 30 minutes at room temperature) is typically recommended as the primary approach because it preserves cellular architecture and is compatible with most antibodies, though it may require optimization of subsequent permeabilization steps.

Alternative approaches include methanol fixation (100% cold (-20°C) methanol for 6-10 minutes), which simultaneously fixes and permeabilizes cells and can expose certain epitopes masked by formaldehyde, though it may distort some cellular structures and extract certain proteins. Glutaraldehyde fixation (0.1-0.5% glutaraldehyde with 3.7% formaldehyde) provides superior structural preservation but can cause higher autofluorescence and may mask epitopes.

Combined approaches might include sequential fixation (short formaldehyde fixation followed by methanol) for improved epitope accessibility, or low concentration dual fixation (2% formaldehyde with 0.2% glutaraldehyde) for balanced preservation. Post-fixation processing is critical, particularly cell wall digestion for S. pombe using zymolyase or lysing enzymes; permeabilization with 0.1% Triton X-100 or 0.5% NP-40 after formaldehyde fixation; and potential autofluorescence reduction using sodium borohydride treatment after glutaraldehyde fixation.

An optimization strategy should test multiple fixation methods in parallel, evaluating based on signal strength, specificity, and morphological preservation, while considering variations in fixation time and temperature.

How does SPAC637.06 compare to homologous proteins in other model organisms?

While detailed information about homologs of SPAC637.06 in other organisms is limited in the available data, researchers can employ several methodological approaches to investigate comparative aspects. For sequence-based homology identification, the UniProt accession number P78817 can be used to retrieve the SPAC637.06 sequence and perform BLAST or HMM-based searches against model organism databases. Both global alignment (full protein) and domain-specific searches should be considered, using tools like Ensembl Compara, OrthoMCL, or InParanoid for comprehensive ortholog detection.

Structural comparison can involve predicting protein structure using tools like AlphaFold or I-TASSER and comparing predicted structures with known structures of potential homologs to identify conserved structural motifs that may indicate functional conservation. Functional domain analysis should identify domains using InterPro, Pfam, or SMART; compare domain architecture with homologs in other organisms; and assess conservation of critical residues within functional domains.

Phylogenetic analysis can construct evolutionary trees using maximum likelihood or Bayesian methods, map functional divergence onto phylogeny, and identify clades with potential functional specialization. Expression pattern comparison between SPAC637.06 and its homologs across conditions may identify conserved regulation patterns indicating functional conservation. For researchers investigating SPAC637.06 homologs, the antibody can be used to validate expression and localization patterns in S. pombe for comparison with homolog localization in other organisms.

What research gaps exist in our understanding of SPAC637.06 function?

Significant research gaps exist in our understanding of SPAC637.06 function, as the protein remains largely uncharacterized. Fundamental functional characterization is lacking, including basic biological role, molecular function, phenotypic consequences of gene deletion or overexpression, and determination of whether the gene is essential or non-essential. Structural information gaps include three-dimensional structure, functional domains and their specific roles, structure-function relationships, and post-translational modification sites and their functional significance.

The protein interaction network remains to be elucidated, including direct binding partners, interaction domains, membership in protein complexes, and dynamic changes in interactions under different conditions. Subcellular localization information is limited, including precise localization pattern, potential dynamic changes during cell cycle or stress, and localization determinants within the protein sequence. Regulatory aspects are unexplored, including transcriptional regulation under different conditions, post-translational modifications and their effects, and protein turnover pathways.

Evolutionary aspects require investigation, including conservation across species beyond S. pombe, functional divergence of homologs, and evolutionary pressure on specific regions. Technical resource limitations include the need for knockout/knockdown strains and tagged versions, and requirements for functional assays specific to SPAC637.06. Integration with known pathways remains to be established, including connections to well-characterized cellular processes. The SPAC637.06 Antibody provides a valuable tool to begin addressing these research gaps, particularly for localization studies, expression analysis, and protein interaction investigations.

How can SPAC637.06 research contribute to broader understanding of eukaryotic cellular processes?

Research on SPAC637.06 can contribute to broader understanding of eukaryotic cellular processes in several significant ways. As S. pombe serves as an excellent model for eukaryotic cell biology with many fundamental processes conserved from yeast to humans, SPAC637.06 research may uncover mechanisms applicable across species. Characterizing SPAC637.06 helps complete the functional annotation of the S. pombe genome, contributing to comprehensive understanding of eukaryotic gene function and addressing the challenge of uncharacterized genes present in all sequenced genomes.

Uncharacterized proteins often represent undiscovered pathways or processes, and SPAC637.06 may function in cellular mechanisms not yet fully understood, potentially revealing new regulatory connections between known pathways. Studying conservation patterns of SPAC637.06 across species can provide evolutionary insights, understanding of functional divergence of homologs, and illuminate the evolution of specific cellular processes.

Given that S. pombe is a premier model for cell cycle research, if SPAC637.06 has cell cycle functions, findings could impact cancer research and may reveal previously unknown aspects of cell division regulation. For translational potential, insights may transfer to human homologs (if they exist) with possible relevance to disease processes in higher eukaryotes. Using tools like the SPAC637.06 Antibody enables researchers to begin systematic characterization, starting with expression patterns and subcellular localization, providing the foundation for understanding this protein's role in eukaryotic cellular biology.