SPBC17D11.03c Antibody

What is SPBC17D11.03c protein and why is it significant for research?

SPBC17D11.03c is a protein from Schizosaccharomyces pombe (fission yeast) with UniProt accession number O74758. This protein is studied in fundamental research on yeast cellular processes. Antibodies targeting this protein allow researchers to investigate its expression, localization, and function within S. pombe, which serves as an important model organism for understanding eukaryotic cell biology. Research with this antibody contributes to our understanding of conserved cellular mechanisms across species .

What detection methods are compatible with SPBC17D11.03c antibody?

SPBC17D11.03c antibody can likely be utilized in multiple experimental techniques similar to other research antibodies. These may include Western blotting, immunoprecipitation, immunohistochemistry, and ELISA. The compatibility depends on the specific antibody formulation and the experimental conditions. When establishing a new application, researchers should perform validation experiments with positive and negative controls to confirm specificity and optimal working conditions, similar to validation processes used for other antibodies like Human IL-17C antibody .

How should SPBC17D11.03c antibody be stored for optimal stability?

Based on standard antibody storage practices, SPBC17D11.03c antibody should likely be stored at 2-8°C for short-term use (up to 12 months from receipt). For applications requiring small aliquots, it's advisable to prepare single-use portions to avoid repeated freeze-thaw cycles. As with other antibodies, protection from light and avoiding freezing may help maintain reactivity and specificity over time .

What controls should be incorporated when using SPBC17D11.03c antibody in Western blot experiments?

For robust Western blot experiments, researchers should include:

• Positive control: Lysate from wild-type S. pombe cells expressing SPBC17D11.03c

• Negative control: Lysate from SPBC17D11.03c knockout/knockdown strain

• Loading control: Antibody against a housekeeping protein like actin

• Isotype control: Similar antibody of the same isotype but targeting an irrelevant protein

These controls help distinguish specific signals from non-specific binding and provide assurance of experimental validity, similar to control strategies used with other research antibodies .

How can researchers optimize immunoprecipitation protocols using SPBC17D11.03c antibody?

For effective immunoprecipitation:

• Determine optimal antibody concentration through titration experiments (typically 2-10 μg per reaction)

• Test different lysis buffers to preserve protein-protein interactions relevant to your experiment

• Consider crosslinking the antibody to beads to prevent antibody co-elution

• Include appropriate negative controls (isotype antibody or pre-immune serum)

• Validate specificity using Western blot analysis of immunoprecipitated material

These approaches resemble methodologies used for immunoprecipitation with other research antibodies, adapted for yeast cell experiments .

What strategies can address weak signal problems when using SPBC17D11.03c antibody in immunofluorescence microscopy?

When experiencing weak signals:

• Optimize fixation method (test aldehyde-based vs. organic solvent fixation)

• Evaluate different cell wall digestion protocols specific for S. pombe

• Increase antibody concentration incrementally (1:500 to 1:100 dilutions)

• Extend primary antibody incubation time (overnight at 4°C)

• Use signal amplification systems (tyramide signal amplification or high-sensitivity detection reagents)

• Adjust image acquisition settings (exposure time, gain)

These approaches help maximize detection while maintaining specificity, similar to optimization strategies used for other antibodies in cellular imaging applications .

How can researchers validate SPBC17D11.03c antibody specificity in their experimental system?

Comprehensive validation includes:

• Comparing staining patterns in wild-type vs. SPBC17D11.03c deletion strains

• Performing peptide competition assays with the immunizing peptide

• Using multiple antibodies targeting different epitopes of the same protein

• Correlating protein detection with mRNA expression data

• Mass spectrometry analysis of immunoprecipitated proteins

These approaches provide multiple lines of evidence for antibody specificity, which is crucial for publication-quality research, following validation principles used for other research antibodies .

What are the considerations for using SPBC17D11.03c antibody in flow cytometry with yeast cells?

For successful flow cytometry with yeast cells:

• Optimize cell fixation and permeabilization protocols specifically for S. pombe cell wall

• Establish appropriate gating strategies based on cell size and granularity

• Test different antibody concentrations to determine optimal signal-to-noise ratio

• Include appropriate isotype controls to establish background fluorescence levels

• Consider using fluorochrome-conjugated secondary antibodies with brightness appropriate for the expected protein abundance

These considerations address the unique challenges of yeast cell analysis by flow cytometry, adapting protocols similar to those used for other intracellular targets .

Does SPBC17D11.03c antibody exhibit cross-reactivity with homologous proteins in other yeast species?

Cross-reactivity analysis requires:

• Sequence alignment of SPBC17D11.03c with potential homologs in related species

• Testing the antibody against lysates from multiple yeast species

• Performing epitope mapping to identify the specific recognition sequence

• Validating specificity through knockout/knockdown experiments in each species

Understanding cross-reactivity is essential for comparative studies across yeast species and ensures accurate interpretation of experimental results .

How can researchers quantitatively compare SPBC17D11.03c protein levels across different experimental conditions?

For quantitative analysis:

• Use standardized lysate preparation methods to ensure consistent extraction efficiency

• Establish a standard curve using recombinant SPBC17D11.03c protein if available

• Implement densitometry analysis for Western blots with appropriate normalization

• Consider ELISA-based quantification for higher throughput experiments

• Use spike-in controls to account for extraction and detection variations

These quantitative approaches enable reliable comparison of protein levels across different conditions or mutant strains, similar to quantification methods used with other research antibodies .

How can SPBC17D11.03c antibody be used in chromatin immunoprecipitation (ChIP) experiments to study protein-DNA interactions?

For successful ChIP applications:

• Optimize crosslinking conditions specifically for S. pombe cells

• Develop sonication protocols that yield DNA fragments of appropriate size (200-500 bp)

• Determine optimal antibody-to-chromatin ratio through titration experiments

• Include appropriate controls (input DNA, IgG control, positive control region)

• Validate ChIP efficiency using qPCR before proceeding to sequencing

These considerations address the specific challenges of performing ChIP in yeast cells and help ensure reliable results for protein-DNA interaction studies .

What approaches can be used to study post-translational modifications of SPBC17D11.03c protein?

To investigate post-translational modifications:

• Use modification-specific antibodies in combination with SPBC17D11.03c antibody

• Perform immunoprecipitation followed by mass spectrometry analysis

• Use phosphatase or deubiquitinase treatments to confirm modification-dependent signals

• Develop site-specific mutants to validate modification sites

• Compare modification patterns across different growth conditions or cell cycle phases

These approaches enable comprehensive characterization of protein regulation through post-translational modifications, similar to methodologies used to study modifications of other proteins .

What considerations are important when using SPBC17D11.03c antibody for co-immunoprecipitation studies to identify protein interaction partners?

For effective co-immunoprecipitation:

• Select lysis conditions that preserve native protein interactions (mild detergents, physiological salt concentration)

• Consider chemical crosslinking to stabilize transient interactions

• Optimize antibody concentration to maximize target protein capture without saturating the system

• Include appropriate negative controls (isotype antibody, unrelated protein antibody)

• Validate interactions through reciprocal co-immunoprecipitation experiments

• Confirm biological relevance through functional studies

These methodological considerations help identify genuine interaction partners while minimizing false positives, similar to co-immunoprecipitation approaches used with other antibodies .

How can researchers develop a quantitative ELISA assay using SPBC17D11.03c antibody?

To establish a quantitative ELISA:

• Determine optimal coating conditions (antibody concentration, buffer, temperature)

• Develop a standard curve using purified recombinant SPBC17D11.03c protein

• Optimize detection antibody (consider using a different clone recognizing a distinct epitope)

• Establish appropriate blocking conditions to minimize background

• Validate assay specificity, sensitivity, and reproducibility

• Determine the linear range and lower limit of detection

These systematic development steps enable creation of a reliable quantitative assay for SPBC17D11.03c protein, following principles used in developing ELISAs for other protein targets .