SPCP1E11.10 Antibody

What is SPCP1E11.10 Antibody and what organism does it target?

SPCP1E11.10 Antibody is a polyclonal antibody raised in rabbits that specifically targets the SPCP1E11.10 protein from Schizosaccharomyces pombe (strain 972 / ATCC 24843), commonly known as fission yeast. This antibody was developed using a recombinant SPCP1E11.10 protein as the immunogen and has been purified via antigen affinity methods .

The target protein (UniProt ID: Q9UU77) is specific to S. pombe, making this antibody a valuable tool for researchers studying this model organism's cellular processes. Unlike antibodies that target human proteins such as SCP1 (synaptonemal complex protein 1) or SCPEP1 (serine carboxypeptidase 1) , this antibody is specifically designed for yeast research applications.

What are the validated applications for SPCP1E11.10 Antibody?

SPCP1E11.10 Antibody has been validated for the following experimental applications:

• Western Blotting (WB): Suitable for protein detection and quantification

• Enzyme-Linked Immunosorbent Assay (ELISA): Effective for sensitive protein detection

These applications have been specifically tested to ensure identification of the antigen with high specificity . Unlike some more broadly applicable antibodies that work across multiple techniques like immunohistochemistry and immunofluorescence, the current validation focuses on protein detection methods.

What are the optimal storage conditions for maintaining SPCP1E11.10 Antibody activity?

For optimal preservation of antibody activity, SPCP1E11.10 Antibody should be stored at -20°C or -80°C immediately upon receipt. Researchers should avoid repeated freeze-thaw cycles as these can significantly degrade antibody performance .

The antibody is supplied in liquid form with a storage buffer containing:

• 0.03% Proclin 300 (preservative)

• 50% Glycerol

• 0.01M PBS (pH 7.4)

This formulation helps maintain stability during storage. The inclusion of glycerol prevents complete freezing at -20°C, reducing damage from ice crystal formation that can occur during freeze-thaw cycles.

How should researchers determine optimal antibody concentrations for Western blot experiments?

When designing Western blot experiments with SPCP1E11.10 Antibody, researchers should:

• Begin with a titration experiment using multiple concentrations (typically 1:500, 1:1000, 1:2000, 1:5000)

• Test against both positive controls (S. pombe extracts) and negative controls (extracts from other organisms)

• Evaluate signal-to-noise ratio for each concentration

• Select the dilution that provides clear specific bands with minimal background

This methodical approach helps establish experimental parameters that maximize specificity while conserving valuable antibody resources. Unlike fixed protocols, antibody concentrations often require optimization for each specific laboratory environment and detection system.

What controls are essential when working with SPCP1E11.10 Antibody?

Proper experimental design with SPCP1E11.10 Antibody requires several controls:

Implementing these controls is critical for producing reliable, reproducible results and properly interpreting experimental outcomes. This approach aligns with rigorous validation practices seen in high-quality antibody characterization, similar to the multi-tiered validation protocols used for clinical antibodies .

How can researchers verify SPCP1E11.10 Antibody specificity in complex S. pombe protein extracts?

For rigorous verification of antibody specificity, researchers should employ multiple complementary approaches:

• Immunoprecipitation followed by mass spectrometry:

  • Perform immunoprecipitation with SPCP1E11.10 Antibody

  • Analyze precipitated proteins by mass spectrometry

  • Confirm presence of SPCP1E11.10 protein and assess co-precipitating proteins

• Genetic validation:

  • Compare antibody reactivity between wild-type and SPCP1E11.10 knockout/knockdown strains

  • Absence of signal in knockout/knockdown samples confirms specificity

• Cross-adsorption experiments:

  • Pre-adsorb antibody with recombinant SPCP1E11.10 protein

  • Reduced or eliminated signal confirms epitope-specific binding

These approaches provide multi-dimensional validation of antibody specificity, crucial for interpreting complex experimental results in advanced research settings.

What methodologies enable quantitative analysis of SPCP1E11.10 expression using this antibody?

Quantitative analysis of SPCP1E11.10 expression requires careful methodological considerations:

• Quantitative Western Blotting:

  • Use fluorescent secondary antibodies rather than chemiluminescence

  • Include calibration curves with known quantities of recombinant protein

  • Employ image analysis software with linear range validation

  • Normalize to validated loading controls

• Quantitative ELISA development:

  • Establish standard curves using purified recombinant SPCP1E11.10

  • Determine lower limit of detection and linear range

  • Validate reproducibility across technical and biological replicates

  • Account for matrix effects from complex biological samples

These methodologies transform qualitative antibody applications into quantitative tools for measuring protein expression levels. The approach shares conceptual similarities with quantitative assays developed for therapeutic antibody assessment in clinical research .

How should researchers address weak or absent signal when using SPCP1E11.10 Antibody?

When facing weak or absent signals, consider this systematic troubleshooting approach:

• Antibody Activity Assessment:

  • Verify storage conditions were maintained

  • Check antibody expiration date

  • Test a new antibody aliquot if possible

• Sample Preparation Optimization:

  • Ensure proper cell lysis to release target protein

  • Consider different extraction buffers to maintain protein antigenicity

  • Verify protein integrity by SDS-PAGE and total protein staining

• Protocol Adjustments:

  • Increase antibody concentration

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

  • Optimize blocking conditions to reduce competition for binding

  • Enhance detection sensitivity using amplification systems

• Expression Verification:

  • Confirm SPCP1E11.10 expression in your specific S. pombe strain and growth conditions

  • Consider RT-PCR to verify mRNA expression before protein analysis

This methodical approach can help identify and resolve common causes of signal problems when working with research antibodies.

What strategies can resolve cross-reactivity issues with SPCP1E11.10 Antibody?

When encountering potential cross-reactivity with non-target proteins:

• Blocking Optimization:

  • Test alternative blocking agents (BSA, casein, commercial blockers)

  • Increase blocking time and concentration

• Washing Modifications:

  • Increase wash buffer stringency (higher salt concentration, addition of detergents)

  • Extend washing times and increase wash steps

• Antibody Dilution Adjustment:

  • Test higher dilutions to reduce non-specific binding

  • Consider using antibody diluent formulations with blocking components

• Pre-adsorption Strategy:

  • Pre-adsorb the antibody with proteins from the species causing cross-reactivity

  • Use acetone powder preparation from non-target tissues for pre-adsorption

• Alternative Detection Methods:

  • If Western blot shows cross-reactivity, try ELISA or vice versa

  • Consider immunoprecipitation followed by Western blotting for higher specificity

These strategies help overcome common cross-reactivity challenges that can complicate the interpretation of experimental results.

How can researchers employ SPCP1E11.10 Antibody in studying protein-protein interactions?

For investigating protein-protein interactions involving SPCP1E11.10:

• Co-immunoprecipitation (Co-IP):

  • Optimize lysis conditions to maintain protein complexes

  • Use gentle elution methods to preserve interactions

  • Confirm results with reciprocal Co-IP using antibodies against suspected interaction partners

  • Consider crosslinking for transient interactions

• Proximity Ligation Assay (PLA) Development:

  • Combine SPCP1E11.10 Antibody with antibodies against potential interacting partners

  • Optimize fixation to preserve protein localization while maintaining epitope accessibility

  • Include appropriate controls to validate interaction specificity

• Pull-down Validation:

  • Use recombinant SPCP1E11.10 protein for pull-down experiments

  • Confirm interactions identified by antibody-based methods

  • Characterize binding domains through truncation studies

These methodologies enable researchers to move beyond simple protein detection and investigate functional protein networks, similar to approaches used in studying complex antibody interactions with target epitopes .

What considerations are important when adapting SPCP1E11.10 Antibody for chromatin immunoprecipitation (ChIP) studies?

Although not currently validated for ChIP applications, researchers interested in adapting SPCP1E11.10 Antibody for chromatin studies should consider:

• Crosslinking Optimization:

  • Test different formaldehyde concentrations (0.1-1%)

  • Evaluate alternative crosslinking agents for protein-DNA interactions

  • Optimize crosslinking time to balance efficiency and reversibility

• Sonication Parameters:

  • Determine optimal sonication conditions to achieve appropriate chromatin fragment size

  • Verify fragment distribution by agarose gel electrophoresis

  • Ensure consistent sonication across samples

• Antibody Evaluation:

  • Perform preliminary ChIP using positive control antibodies (e.g., histone modifications)

  • Test SPCP1E11.10 Antibody at various concentrations

  • Compare ChIP efficiency to Western blot signal intensity

• Controls Implementation:

  • Include input chromatin controls

  • Use non-specific IgG as negative control

  • Consider spike-in controls for normalization

• Signal Validation:

  • Confirm enrichment by qPCR at expected genomic regions

  • Validate findings with alternative methods (e.g., reporter assays)

This methodical approach allows researchers to explore potential chromatin associations of SPCP1E11.10, extending its research applications beyond conventional protein detection methods.

How does SPCP1E11.10 Antibody compare to other fission yeast antibodies in specificity and application range?

When evaluating SPCP1E11.10 Antibody against other S. pombe antibodies:

• Specificity Comparison:

  • SPCP1E11.10 Antibody is highly specific for its target protein in S. pombe

  • Unlike broader spectrum antibodies that recognize conserved proteins across species, this antibody offers high specificity for strain-specific research

  • The polyclonal nature may provide recognition of multiple epitopes, potentially increasing sensitivity compared to monoclonal alternatives

• Application Range:

  • Currently validated for ELISA and Western blotting

  • More specialized than antibodies targeting conserved cellular components (e.g., actin, tubulin)

  • May require additional validation for applications beyond those currently verified

• Research Context:

  • Particularly valuable for studies focused on SPCP1E11.10 protein function in S. pombe

  • May be less suitable for cross-species comparative studies due to high specificity

  • Offers advantages in studies requiring selective protein detection in complex S. pombe extracts

This comparative perspective helps researchers select the most appropriate antibody for their specific research objectives and experimental systems.

What emerging methodologies could expand the utility of SPCP1E11.10 Antibody in fission yeast research?

Researchers should consider these emerging approaches:

• Single-cell Protein Analysis:

  • Adapting SPCP1E11.10 Antibody for flow cytometry or mass cytometry

  • Developing protocols for in situ protein detection in individual yeast cells

  • Combining with DNA content analysis for cell-cycle studies

• Spatiotemporal Dynamics:

  • Optimizing for live-cell imaging applications through recombinant antibody fragments

  • Developing antibody-based biosensors for real-time protein dynamics

  • Combining with optogenetic approaches for manipulating protein function

• Multi-omic Integration:

  • Using antibody-based pull-downs for integrated proteomics and genomics

  • Developing ChIP-seq protocols to map genomic interactions

  • Correlation of protein levels with transcriptomic data for systems biology approaches

These forward-looking applications represent the frontier of research antibody utilization, similar to advances seen in therapeutic antibody development .