RPS11A Antibody

What is RPS11 and what is its biological significance?

RPS11 (40S ribosomal protein S11) is a component of the 40S ribosomal subunit, which together with the 60S subunit forms the complete ribosome that catalyzes protein synthesis. It belongs to the S17P family of ribosomal proteins. The gene encoding RPS11 is co-transcribed with the small nucleolar RNA gene U35B, which is located in the third intron . As with many ribosomal proteins, multiple processed pseudogenes of RPS11 are dispersed throughout the genome. RPS11 plays a crucial role in the fundamental cellular process of protein translation, making it essential for normal cellular function and viability.

Beyond its canonical role in translation, research has revealed that RPS11 may have additional functions, particularly in cancer biology. Studies have shown elevated expression of RPS11 in various malignancies, suggesting potential roles in carcinogenesis and tumor progression .

What are the typical applications for RPS11 antibodies in research?

RPS11 antibodies have several important applications in research:

• Western Blotting (WB): Used at dilutions of 1:500-1:2000 to detect RPS11 protein in cell and tissue lysates. This technique allows for the quantification of RPS11 expression levels and comparison between different experimental conditions or sample types .

• Immunohistochemistry (IHC): Applied at dilutions of 1:50-1:200 for detecting RPS11 in tissue sections. This technique is valuable for examining RPS11 expression patterns in different cell types within tissues, particularly in cancer samples .

• ELISA: Typically used at dilutions around 1:5000 for quantitative detection of RPS11 in solution .

• Prognostic Marker Analysis: Used in clinical research to evaluate RPS11 as a potential biomarker for patient outcomes, particularly in hepatocellular carcinoma and other cancers .

What are the recommended storage conditions for RPS11 antibodies?

For optimal retention of activity, RPS11 antibodies should be stored at -20°C, where they remain valid for approximately 12 months . The antibodies are typically shipped at 4°C, but upon receipt, they should immediately be stored at the recommended temperature of -20°C . To maintain antibody integrity, it's crucial to avoid repeated freeze/thaw cycles, as these can lead to protein denaturation and loss of binding activity .

Most commercial RPS11 antibodies are supplied in a stabilized formulation containing phosphate buffered saline (pH 7.4) with sodium azide (0.02-0.05%) as a preservative and 50% glycerol to prevent freezing at -20°C and maintain stability .

How can RPS11 antibodies be used to study hepatocellular carcinoma?

RPS11 antibodies have proven valuable in hepatocellular carcinoma (HCC) research through several methodological approaches:

What is the relationship between RPS11 expression and cancer beyond HCC?

Research has identified altered RPS11 expression across multiple cancer types:

This consistent pattern of RPS11 overexpression across diverse malignancies suggests a potential universal role in carcinogenesis, making it an important target for cancer research beyond just HCC.

What biological pathways might be associated with RPS11 in cancer research?

While the complete picture of RPS11's role in cancer biology continues to emerge, research has begun to identify potential mechanisms and pathways:

• Pathway Analysis: Single-sample gene-set enrichment analysis (ssGSEA) in TCGA liver cancer databases has been used to investigate biological pathways associated with RPS11 expression .

• Extra-Ribosomal Functions: Beyond its canonical role in protein synthesis, emerging evidence suggests RPS11 may have non-ribosomal functions that influence cancer progression, similar to other ribosomal proteins that have been shown to regulate cell cycle progression and p53 signaling.

• Association with Tumor Characteristics: The correlation between high RPS11 expression and poor tumor differentiation in HCC suggests potential involvement in pathways governing cellular differentiation and malignant transformation .

Understanding these pathways will require further investigation using RPS11 antibodies in conjunction with other research tools to elucidate the precise mechanisms by which RPS11 contributes to cancer development and progression.

What are the optimal conditions for Western blotting with RPS11 antibodies?

For successful Western blotting with RPS11 antibodies, researchers should consider the following protocol parameters:

What considerations are important for immunohistochemistry with RPS11 antibodies?

When performing immunohistochemistry with RPS11 antibodies, researchers should consider these methodological details:

• Tissue Preparation: Formalin-fixed, paraffin-embedded tissue sections should be subjected to appropriate antigen retrieval. In published protocols, tissue microarrays were treated with hydrogen peroxide (3% H₂O₂) solution at room temperature for 15 minutes to block endogenous peroxidase activity .

• Antibody Dilution and Incubation: The recommended dilution range is 1:50-1:200 . Optimal results have been reported when sections are incubated with RPS11 antibody (1:100, e.g., Abcam, Cat No# ab175213) overnight at 4°C .

• Detection System: After primary antibody incubation, sections should be treated with horseradish peroxidase (HRP)-conjugated secondary antibody for 45 minutes at 37°C, followed by diaminobenzidine (DAB) solution for visualization .

• Assessment Method: The H-score method provides quantitative evaluation of RPS11 IHC staining by multiplying staining intensity (negative: 0, weak: 1, moderate: 2, strong: 3) by staining extent (0–100%) .

• Verified Tissues: Human tonsil and human prostate cancer tissues have been verified for successful RPS11 detection by IHC .

How can researchers validate the specificity of RPS11 antibodies?

Ensuring antibody specificity is crucial for reliable research results. Several validation approaches are recommended:

• Western Blot Profile: Confirm that the antibody detects a band of the expected molecular weight (approximately 18 kDa) in appropriate positive control samples. Note that observed band sizes may vary due to post-translational modifications .

• Blocking Peptide Controls: Perform parallel experiments where the antibody is pre-incubated with the immunizing peptide (derived from human RPS11 amino acids 71-120 in some commercial antibodies). This should abolish or significantly reduce the specific signal .

• Knockdown/Knockout Controls: When possible, validate antibody specificity using siRNA knockdown or CRISPR/Cas9 knockout of RPS11, which should result in reduction or elimination of the specific signal.

• Cross-Reactivity Assessment: If working across species, confirm the antibody's reactivity with the target species. Many commercial RPS11 antibodies are verified for human, mouse, and rat samples .

• Multiple Antibody Comparison: When feasible, compare results using different antibodies that recognize different epitopes of RPS11 to increase confidence in the specificity of detection.

What evidence exists for RPS11 as a prognostic biomarker in hepatocellular carcinoma?

Substantial evidence supports RPS11's potential as a prognostic biomarker in hepatocellular carcinoma (HCC):

These consistent findings across multiple research approaches and patient cohorts strongly suggest that RPS11 could serve as a valuable prognostic biomarker in HCC.

How does RPS11 expression correlate with clinical parameters in HCC patients?

Research has identified several significant correlations between RPS11 expression and clinical parameters in HCC patients:

These correlations were consistent across both training (n=182) and validation (n=90) cohorts of HCC patients . The strong association with established HCC biomarkers like AFP reinforces the potential clinical relevance of RPS11 expression assessment in HCC management.

Other clinical parameters, including age, gender, HBsAg status, and CEA levels, did not show significant associations with RPS11 expression levels .