rps1201 Antibody
Description
Definition and Target
The rps1201 Antibody is a monoclonal antibody targeting the ribosomal protein S12 (RPS12) in Schizosaccharomyces pombe (fission yeast). Ribosomal proteins like RPS12 are critical for translational accuracy and ribosome assembly. This antibody is cataloged under the product code CSB-PA522629XA01SXV and corresponds to UniProt ID O14062 .
Biological Context of RPS12
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Role in Translation: RPS12 is a component of the 30S ribosomal subunit, working alongside S4 and S5 to ensure translational fidelity .
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Conservation: Homologs of RPS12 exist across eukaryotes, with functional studies in yeast providing insights into ribosome structure and function .
Research Applications
While specific studies using rps1201 Antibody are not detailed in the provided sources, its applications likely include:
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Localization Studies: Mapping RPS12 distribution in fission yeast cells via immunofluorescence.
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Protein Interaction Analysis: Identifying RPS12-associated complexes using immunoprecipitation.
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Functional Knockdown Experiments: Validating RPS12’s role in ribosome assembly or stress responses.
Table 2: Antibodies Targeting Ribosomal Proteins in S. pombe
| Antibody Name | Target Protein | UniProt ID | Applications |
|---|---|---|---|
| rps1201 | RPS12 | O14062 | WB, IF, IP |
| rps3 | RPS3 | O60128 | WB, ELISA |
| rps2201 | RPS22 | P0CT58 | Immunohistochemistry |
Source: Cusabio product catalog
Limitations and Future Directions
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Species Specificity: This antibody is validated for fission yeast and may not cross-react with human or other eukaryotic RPS12 homologs.
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Research Gaps: Published studies directly utilizing rps1201 Antibody are absent in the reviewed literature, highlighting opportunities for novel investigations into ribosomal biology.
References
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Q&A
What validated applications are available for RPS12 antibody (16490-1-AP)?
The RPS12 antibody has been experimentally validated for the following applications, with specific performance metrics (Table 1):
For IHC, antigen retrieval is critical to unmask epitopes in formalin-fixed tissues. The choice of buffer (TE vs. citrate) may influence background noise and signal-to-noise ratio, requiring empirical testing .
How should antibody dilution be optimized for different applications?
Optimal dilution depends on experimental conditions (Table 2):
Methodological Approach:
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Pilot Experiments: Test 2–3 dilutions within the recommended range.
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Signal-to-Noise Ratio: Prioritize dilutions that maximize target band intensity while minimizing background.
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Cross-Validation: Compare results across cell lines or tissues to confirm consistency .
What species does this antibody target, and how to assess cross-reactivity?
The antibody is validated for human, mouse, and rat samples, with cited reactivity in Drosophila in one study . For untested species:
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Western Blot: Use lysates from target species to detect the 15 kDa band.
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Control Experiments: Include lysates from species with known reactivity (e.g., human HeLa cells) as positive controls.
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Sequence Alignment: Compare RPS12 protein sequences between species to predict epitope conservation .
What protocols are recommended for specific applications?
Protocols are available for WB, IHC, IF, and IP, with critical parameters:
Downloadable Protocols: Links are provided for streamlined adoption .
How to troubleshoot no signal in IHC?
Common Causes and Solutions:
What is the molecular weight of RPS12 detected by this antibody?
The antibody detects a 15 kDa protein, consistent with both calculated (132 aa) and observed molecular weights . This ensures specificity for RPS12 (ribosomal protein S12) and avoids cross-reactivity with larger homologs.
How to validate antibody specificity in knockdown/knockout (KD/KO) models?
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KD/KO Cell Validation: Use CRISPR-edited RPS12-KO cells as negative controls.
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Western Blot: Compare signal intensity between wild-type and KO lysates.
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Published Validation: Reference studies where this antibody was used in KD/KO contexts (e.g., ribosome biogenesis studies) .
What storage conditions preserve antibody activity?
Store at -20°C in PBS with 0.02% sodium azide and 50% glycerol. Avoid freeze-thaw cycles; aliquoting is unnecessary for long-term storage .
How to interpret variable band intensity across cell lines in WB?
Factors Influencing Signal:
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Expression Levels: RPS12 abundance varies by cell type (e.g., higher in rapidly dividing cells).
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Sample Preparation: Lysis efficiency and protein concentration.
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Experimental Conditions: Loading quantity (e.g., 20–50 µg total protein per lane).
Recommendation: Normalize to housekeeping proteins (e.g., β-actin) and include technical replicates .
What advanced applications have been reported for this antibody?
Published Uses:
Emerging Methods: Combining RPS12 detection with ribosome profiling to study translational regulation .
