SPAC11D3.17 Antibody

What is SPAC11D3.17 antibody and what organism does it target?

SPAC11D3.17 antibody is a polyclonal antibody raised in rabbits against a recombinant Schizosaccharomyces pombe (strain 972/ATCC 24843) SPAC11D3.17 protein. It specifically targets proteins in yeast species and is available as an unconjugated antibody for research applications . The antibody has been affinity-purified and is supplied with both positive control antigens (200μg) and negative control pre-immune serum (1ml), which are essential components for proper experimental validation .

What applications is the SPAC11D3.17 antibody validated for?

According to the product information, SPAC11D3.17 antibody has been validated for two primary applications:

It's crucial to note that validation in these applications does not automatically extend to other experimental techniques. Researchers must perform additional validation if planning to use the antibody in non-validated applications .

How should SPAC11D3.17 antibody be stored to maintain its activity?

For optimal preservation of antibody activity, SPAC11D3.17 antibody should be stored at either -20°C or -80°C according to the manufacturer's specifications . Repeated freeze-thaw cycles should be avoided as they can lead to antibody degradation and reduced performance. For working solutions, aliquoting the antibody upon first thaw is recommended to minimize freeze-thaw cycles and maintain consistent performance across experiments .

What validation methods should be applied before using SPAC11D3.17 antibody in research?

Following the "five pillars" of antibody characterization established by the International Working Group for Antibody Validation, researchers should implement at least one of these validation strategies for SPAC11D3.17 antibody:

• Genetic strategies: Using knockout or knockdown techniques to confirm specificity.

• Orthogonal strategies: Comparing antibody-dependent results with antibody-independent methods.

• Multiple independent antibody strategies: Using different antibodies targeting the same protein to verify results.

• Recombinant expression strategies: Increasing target protein expression to confirm detection.

• Immunocapture MS strategies: Using mass spectrometry to identify proteins captured by the antibody .

The validation approach should be selected based on experimental context and available resources, with thorough documentation of validation results .

What controls should be included when using SPAC11D3.17 antibody in experiments?

A comprehensive control strategy for SPAC11D3.17 antibody experiments should include:

• Positive controls: Using the provided 200μg of antigens to confirm antibody activity.

• Negative controls: Using the provided pre-immune serum to assess non-specific binding.

• Technical controls: Including no-primary-antibody controls to evaluate secondary antibody specificity.

• Biological controls: When possible, using samples with known expression levels of the target protein, including samples where the target is absent.

These controls help distinguish true signals from background noise and confirm specificity of the observed results .

How can I determine if my batch of SPAC11D3.17 antibody maintains the expected specificity?

Batch-to-batch variability is a significant concern, particularly with polyclonal antibodies like SPAC11D3.17. To verify specificity with each new batch:

• Perform side-by-side comparison with a previously validated batch using the same experimental conditions.

• Compare specific binding patterns in known positive and negative samples.

• Document the batch number in your experimental records and publications.

• Consider testing the antibody against recombinant SPAC11D3.17 protein at varying concentrations to establish a detection threshold.

This systematic approach helps identify potential variability that could impact experimental results and reproducibility .

What factors should be considered when optimizing SPAC11D3.17 antibody for Western blot applications?

Optimizing SPAC11D3.17 antibody for Western blot requires systematic adjustment of multiple parameters:

• Antibody concentration: Begin with the manufacturer's recommended dilution (for similar antibodies, typically 1:2000 for Western blot) and adjust as needed.

• Sample preparation: Ensure proper cell lysis and protein denaturation protocols compatible with the yeast species.

• Blocking conditions: Test different blocking agents (BSA, milk, commercial blockers) to minimize background.

• Incubation time and temperature: Compare short incubations at room temperature versus overnight at 4°C.

• Detection system: Select appropriate secondary antibody and detection method based on expected abundance of target protein.

Document all optimization steps methodically for reproducibility and consider validating the optimized protocol with the positive control antigen provided with the antibody .

How should experimental data using SPAC11D3.17 antibody be reported in publications?

Comprehensive reporting of SPAC11D3.17 antibody usage should include:

• Complete antibody identification: Manufacturer (Cusabio), catalog number (CSB-PA605934XA01SXV-2), clone type (polyclonal), host species (rabbit).

• Experimental application: Detailed methods for each application (ELISA, Western blot).

• Validation evidence: Description of validation performed or citation of previous validation.

• Batch information: Batch or lot number to account for potential batch variation.

• Dilution and conditions: Specific working concentration and experimental conditions.

This detailed reporting enables proper evaluation of research findings and supports experimental reproducibility by other researchers .

What are common causes of non-specific binding with SPAC11D3.17 antibody and how can they be addressed?

Non-specific binding can manifest as multiple bands in Western blots or diffuse signals in other applications. Common causes and solutions include:

• Insufficient blocking: Increase blocking time or test alternative blocking agents.

• Cross-reactivity: Perform pre-adsorption with related proteins or use more stringent washing conditions.

• Excessive antibody concentration: Titrate to determine optimal concentration that maximizes specific signal while minimizing background.

• Sample complexity: Consider pre-clearing complex samples or implementing additional purification steps.

• Detection system sensitivity: Adjust exposure time or switch to less sensitive detection methods if background is excessive.

When troubleshooting, change only one variable at a time and document all modifications to the protocol .

How can I distinguish between true binding and artifacts when using SPAC11D3.17 antibody?

To differentiate genuine signals from artifacts:

• Validate with orthogonal methods: Confirm results using techniques that don't rely on antibodies, such as mass spectrometry.

• Competitor assays: Demonstrate signal reduction when adding excess target protein.

• Specificity controls: Use the pre-immune serum (provided) as a negative control under identical conditions.

• Expected molecular weight: Verify that observed bands match the predicted molecular weight of SPAC11D3.17 protein.

• Reproducibility testing: Confirm consistent results across multiple experimental replicates.

This multi-faceted approach provides stronger evidence for genuine binding versus artifactual signals .

How can computational modeling inform the use of SPAC11D3.17 antibody in complex experimental designs?

Advanced computational approaches can enhance SPAC11D3.17 antibody applications:

• Specificity prediction: Biophysics-informed models can predict potential cross-reactivity with related proteins.

• Epitope mapping: Computational prediction of antibody binding sites can help understand potential cross-reactivity.

• Binding mode analysis: Models can identify distinct binding modes associated with specific or non-specific interactions.

• Optimization algorithms: Computational methods can guide experimental design by predicting optimal conditions.

These computational tools can complement experimental validation and guide the development of highly specific protocols, particularly when working with complex yeast protein mixtures .

What approaches can be used to enhance the specificity of SPAC11D3.17 antibody for challenging applications?

For applications requiring enhanced specificity:

• Pre-adsorption: Incubating the antibody with related proteins to remove cross-reactive antibodies.

• Affinity purification: Further purifying the antibody against the target antigen.

• Competitive binding assays: Including soluble target protein to compete for non-specific binding.

• Sequential immunoprecipitation: Performing multiple rounds of immunocapture to increase specificity.

• Combined approach validation: Implementing multiple validation methods from the "five pillars" to confirm specificity in challenging contexts.

These approaches are particularly valuable when working with closely related proteins or in applications where cross-reactivity poses significant challenges to data interpretation .

What considerations are important when using SPAC11D3.17 antibody in multi-omics or integrative research approaches?

In multi-omics research contexts:

• Cross-platform validation: Verify findings across proteomic, transcriptomic, and genomic datasets.

• Sample preparation compatibility: Ensure antibody performance is maintained in sample preparation methods compatible with multi-omics workflows.

• Data integration strategies: Develop analytical frameworks that account for the limitations and strengths of antibody-based detection.

• Statistical approaches: Implement appropriate statistical methods for integrating antibody-derived data with other omics datasets.

• Validation hierarchy: Establish which platform provides the gold standard for validation of contradictory results.

This integrated approach strengthens research findings and provides a more comprehensive understanding of biological systems while accounting for the specific characteristics of antibody-based detection methods .