SPCC1020.13c Antibody

What are the basic characteristics of the SPCC1020.13c Antibody?

The SPCC1020.13c Antibody is a polyclonal antibody raised in rabbit against recombinant Schizosaccharomyces pombe (strain 972/ATCC 24843) SPCC1020.13c protein . Key specifications include:

What techniques can SPCC1020.13c Antibody be used for?

Based on the product information, SPCC1020.13c Antibody has been validated for use in ELISA (Enzyme-Linked Immunosorbent Assay) and Western Blot (WB) applications . These techniques are fundamental for detecting and quantifying specific proteins in complex biological samples. The antibody could potentially be used in:

• Protein expression studies in S. pombe

• Detection of post-translational modifications

• Localization studies (though immunofluorescence applications would need validation)

• Characterization of protein-protein interactions (with additional validation)

How does antibody validation differ between monoclonal and polyclonal antibodies like SPCC1020.13c?

Antibody validation approaches differ significantly between monoclonal and polyclonal antibodies. For a polyclonal antibody like SPCC1020.13c, validation should include:

• Specificity testing: Unlike monoclonals, polyclonal antibodies contain a mixture of antibodies recognizing different epitopes on the target antigen. For SPCC1020.13c Antibody, specificity should be validated using:

  • Western blots with positive controls (recombinant SPCC1020.13c protein)

  • Negative controls (lysates from other species)

  • Peptide competition assays

• Cross-reactivity assessment: The antibody documentation mentions species reactivity specific to S. pombe , but researchers should verify this experimentally, especially when the antibody might be used in comparative studies.

• Lot-to-lot consistency: Polyclonal antibodies can exhibit batch variation, so researchers should establish internal validation protocols for each new lot .

What strategies can be employed for optimizing SPCC1020.13c Antibody usage in experimental protocols?

Optimization strategies for SPCC1020.13c Antibody should account for its specific characteristics:

• Titration experiments: Determine optimal antibody concentration for each application by testing a range (e.g., 0.1-10 μg/mL for ELISA, 1:500-1:5000 dilution for Western blot).

• Buffer optimization: While the storage buffer is specified (PBS with glycerol and preservative) , experimental buffers may need optimization:

  • For Western blot: Test different blocking agents (5% BSA vs. non-fat milk)

  • For ELISA: Optimize coating buffer pH and washing stringency

• Incubation conditions: Optimize time (1-24 hours) and temperature (4°C, room temperature).

• Signal enhancement: For low-abundance proteins, consider signal amplification methods such as:

  • Enhanced chemiluminescence for Western blots

  • Biotin-streptavidin systems for ELISA

  • Rolling circle amplification methods (as described in for other antibody applications)

What are the recommended protocols for using SPCC1020.13c Antibody in Western blotting?

Based on general antibody protocols and the specific properties of SPCC1020.13c Antibody, a recommended Western blot protocol would include:

• Sample preparation:

  • Lyse S. pombe cells in appropriate buffer containing protease inhibitors

  • Determine protein concentration (Bradford or BCA assay)

  • Denature samples in reducing Laemmli buffer at 95°C for 5 minutes

• Gel electrophoresis and transfer:

  • Load 20-40 μg total protein per lane

  • Use 10-12% SDS-PAGE for separation

  • Transfer to PVDF membrane (recommended for most antibody applications)

• Immunoblotting:

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

  • Incubate with SPCC1020.13c Antibody at 1:1000 dilution in blocking buffer overnight at 4°C

  • Wash 3-5 times with TBST

  • Incubate with HRP-conjugated anti-rabbit secondary antibody at 1:5000 dilution for 1 hour

  • Develop using ECL substrate and appropriate imaging system

• Controls:

  • Positive control: Recombinant SPCC1020.13c protein

  • Negative control: Non-transformed yeast lysate or lysate from a SPCC1020.13c knockout strain

What approaches should be used to troubleshoot non-specific binding or weak signals when using SPCC1020.13c Antibody?

When encountering issues with SPCC1020.13c Antibody in experiments, systematic troubleshooting approaches include:

For non-specific binding:

• Increase blocking stringency (5% to 10% blocking agent)

• Add 0.1-0.3% Tween-20 to antibody dilution buffer

• Pre-adsorb the antibody with lysates from species that show cross-reactivity

• Increase washing duration and number of wash steps

• Titrate antibody to find optimal concentration that minimizes background

For weak signals:

• Increase antibody concentration or incubation time

• Reduce washing stringency slightly

• Use more sensitive detection systems (enhanced chemiluminescence)

• Enrich for the target protein using subcellular fractionation

• Confirm target protein expression levels using RT-PCR or other methods

How can researchers verify the specificity of SPCC1020.13c Antibody for their particular experimental system?

Verifying antibody specificity is critical for reliable research outcomes. For SPCC1020.13c Antibody, researchers should:

• Perform genetic validation using:

  • SPCC1020.13c knockout/knockdown strains as negative controls

  • SPCC1020.13c overexpression systems as positive controls

• Conduct epitope mapping to determine the specific region recognized by the antibody, which can help predict potential cross-reactivity.

• Implement orthogonal detection methods such as:

  • Mass spectrometry identification of immunoprecipitated proteins

  • RNA expression correlation with protein detection levels

  • Alternative antibodies targeting different epitopes of the same protein

• Document antibody performance metrics:

  • Limit of detection

  • Dynamic range

  • Reproducibility across experiments

These approaches align with best practices in antibody validation as described in the literature for other antibody applications .

What considerations should researchers keep in mind when interpreting results from experiments using SPCC1020.13c Antibody?

When interpreting results from experiments using SPCC1020.13c Antibody, researchers should consider:

• Technical limitations:

  • Polyclonal nature may lead to batch-to-batch variation

  • Possible cross-reactivity with structurally similar proteins

  • Sensitivity limitations in detecting low-abundance proteins

• Biological context:

  • Expression levels of SPCC1020.13c may vary under different conditions

  • Post-translational modifications might affect antibody recognition

  • Protein localization and accessibility could impact detection efficiency

• Experimental design factors:

  • Sample preparation methods may affect epitope availability

  • Buffer conditions can influence antibody-antigen interactions

  • Fixation methods (for immunofluorescence) might impact epitope recognition

• Alternative confirmation approaches:

  • Correlate results with mRNA expression data

  • Validate findings using alternative detection methods

  • Consider using tagged protein expression for orthogonal validation

How might SPCC1020.13c Antibody be used in studies investigating yeast cell biology and evolution?

SPCC1020.13c Antibody could serve as a valuable tool in comparative biology studies:

• Evolutionary conservation analyses:

  • Investigate functional conservation of SPCC1020.13c homologs across yeast species

  • Study structural similarities with related proteins in higher eukaryotes

  • Examine changes in expression patterns across evolutionary lineages

• Cell cycle and stress response studies:

  • Monitor SPCC1020.13c expression during different cell cycle phases

  • Investigate protein regulation under various stress conditions

  • Examine potential roles in cell signaling pathways

• Protein-protein interaction networks:

  • Use in co-immunoprecipitation studies to identify interaction partners

  • Investigate the protein's role in specific cellular complexes

  • Map functional relationships within molecular networks

Similar approaches have been used successfully in studies of other antibodies against yeast proteins, revealing important insights into cellular processes and disease mechanisms .

What are the approaches for integrating SPCC1020.13c Antibody data with other -omics datasets for systems biology studies?

Integrating antibody-based protein detection with other -omics approaches can provide comprehensive insights:

• Multi-omics integration strategies:

  • Correlate protein expression (detected by SPCC1020.13c Antibody) with transcriptomics data

  • Combine with metabolomics to link protein function to metabolic pathways

  • Integrate with phosphoproteomics to study post-translational regulation

• Network biology approaches:

  • Map protein-protein interactions using antibody-based pull-downs

  • Connect to STRING database entries for SPCC1020.13c

  • Develop functional interaction networks across biological scales

• Computational modeling:

  • Use antibody-derived protein quantification for parameter estimation in systems models

  • Incorporate protein level data into predictive models of cellular behavior

  • Validate computational predictions with experimental antibody-based studies

These approaches align with modern systems biology frameworks that have been successfully applied in other antibody-based research contexts .