CRRSP10 Antibody
Description
Functional Context
While specific functional studies on CRRSP10 are not detailed in the provided sources, its nomenclature ("CRRSP") suggests a potential role in cell wall-related processes or stress responses based on analogous Arabidopsis proteins. Other entries in the same database (e.g., "cell wall" antibodies targeting 36 kDa, 48 kDa, and 60 kDa proteins) imply CRRSP10 may participate in structural or regulatory pathways within plant cells .
Applications in Research
The antibody is primarily used for:
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Immunodetection: Western blot (WB), immunohistochemistry (IHC), or ELISA to localize CRRSP10 in Arabidopsis tissues.
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Functional Studies: Investigating CRRSP10's role in developmental or stress-response pathways.
Validation and Quality Control
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Specificity Testing: Via knockout/knockdown models or peptide-blocking assays.
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Batch Consistency: Quality checks for lot-to-lot reproducibility .
Research Gaps and Future Directions
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Structural Characterization: The CRRSP10 protein’s 3D structure and interaction partners remain unstudied.
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In Vivo Role: No peer-reviewed studies directly link CRRSP10 to specific biological pathways.
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Cross-Reactivity: Testing in related plant species (e.g., Brassica napus) could expand utility.
Comparative Analysis with Other Antibodies
The following table contrasts CRRSP10 with select Arabidopsis antibodies from the same source:
| Antibody Target | Product Code | UniProt No. | Key Features |
|---|---|---|---|
| CRRSP10 | CSB-PA865894XA01DOA | Q9SIP6 | Cell wall-associated (inferred) |
| CRRSP8 | CSB-PA882853XA01DOA | Q9SH40 | Hypothetical protein |
| CRRSP17 | CSB-PA885411XA01DOA | Q9LRM2 | Expressed in root tissues |
Authoritative Recommendations
To ensure reliable results with the CRRSP10 Antibody:
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Optimize Dilutions: Conduct checkerboard assays for WB/IHC.
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Include Controls: Use Arabidopsis mutants lacking CRRSP10 (if available) to confirm specificity.
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Collaborate: Share findings via open platforms like the Structural Genomics Consortium to enhance reproducibility .
Product Specs
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
KEGG: ath:AT2G31620
UniGene: At.52996
Q&A
FAQs for CRRSP10 Antibody in Academic Research
What experimental strategies are recommended for validating CRRSP10 antibody specificity in Arabidopsis thaliana?
Validation should include:
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Western Blot (WB): Use recombinant CRRSP10 protein (UniProt: Q9SIP6) as a positive control and knockout Arabidopsis lines as negative controls .
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ELISA: Test antibody-antigen interaction using synthetic peptides representing N-terminal, C-terminal, or internal epitopes (e.g., X-Q9SIP6-N/C/M combinations) .
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Cross-reactivity screening: Assess against homologs in closely related plant species (e.g., Brassica napus) to confirm species specificity .
Table 1: CRRSP10 Antibody Validation Workflow
| Step | Method | Purpose | Key Controls |
|---|---|---|---|
| 1 | WB | Confirm target protein detection | Recombinant CRRSP10, knockout lysate |
| 2 | ELISA | Quantify binding affinity | Peptide competitors (e.g., N-terminal vs. C-terminal) |
| 3 | Immunoprecipitation | Assess functional utility | Isotype-matched antibodies |
How do polyclonal and monoclonal CRRSP10 antibodies differ in research applications?
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Polyclonal antibodies (e.g., CSB-PA865894XA01DOA ):
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Advantages: Detect multiple epitopes, useful for denatured proteins in WB.
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Limitations: Batch-to-batch variability; require rigorous cross-adsorption to minimize off-target binding.
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Monoclonal antibody combinations (e.g., X-Q9SIP6-N/C/M ):
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Advantages: Epitope-specific, reproducible across experiments.
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Applications: Epitope mapping, structural studies, or distinguishing CRRSP10 isoforms.
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How can researchers resolve contradictions in CRRSP10 localization studies?
Conflicting results (e.g., secretory vs. cytoplasmic localization) may arise from:
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Antibody epitope accessibility: Antibodies targeting the N-terminus (X-Q9SIP6-N) may fail to detect post-translationally modified forms .
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Fixation artifacts: Optimize permeabilization protocols for immunolocalization .
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Validation: Combine multiple antibodies (e.g., N- and C-terminal combinations) and orthogonal methods (e.g., CRISPR-Cas9 tagging) .
What computational and experimental approaches improve epitope mapping for CRRSP10?
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Active learning frameworks: Prioritize high-impact antigen-antibody pairs for testing, reducing experimental costs by 35% .
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Structural prediction tools: Use AlphaFold2 to model CRRSP10-antibody complexes and guide mutagenesis studies .
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Peptide arrays: Screen overlapping 15-mer peptides to identify linear epitopes .
Table 2: Epitope Mapping Strategies
| Method | Resolution | Throughput | Cost |
|---|---|---|---|
| Cryo-EM | Atomic (3.1 Å) | Low | High |
| Peptide array | Linear epitopes | High | Moderate |
| HDX-MS | Conformational epitopes | Medium | High |
How can researchers address reproducibility challenges in CRRSP10 studies?
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Antibody characterization: Follow RRID standards and report validation data (e.g., lot numbers, dilution factors) .
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Data sharing: Deposit raw WB/ELISA images in public repositories (e.g., Zenodo).
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Negative controls: Include knockout lines and irrelevant isotype antibodies in every experiment .
What are optimal protocols for CRRSP10 functional studies in plant stress responses?
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Knockdown/out models: Use CRISPR-Cas9 to generate crrsp10 mutants and complementation lines.
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Phenotypic assays: Monitor ion transport or pathogen resistance under abiotic stress.
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Interaction screens: Perform co-IP/MS with CRRSP10 antibodies to identify binding partners .
Methodological Considerations
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Antibody dilution optimization: For WB, test 1:500–1:5,000 dilutions; for ELISA, use 1:10,000 .
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Batch validation: Revalidate new antibody lots using standardized lysates .
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Multiplexing: Pair CRRSP10 antibodies with organelle-specific markers (e.g., ER-Tracker) for colocalization studies.
