MRPL13 Antibody

What is MRPL13 and what is its function in normal cellular biology?

MRPL13, also known as 39S ribosomal protein L13 mitochondrial or L13mt, is a 178 amino acid protein belonging to the ribosomal protein L13P family . It localizes in the mitochondrion and plays a crucial role in synthesizing mitochondrial proteins in cells . As a member of the mitochondrial ribosomal protein family, MRPL13 is integral to the translation of mitochondrial mRNAs, which is essential for proper mitochondrial function and cellular energy production.

The protein has a calculated molecular weight of 21 kDa, though the observed molecular weight in experimental systems typically ranges from 20-23 kDa . The gene encoding MRPL13 in humans has the GenBank Accession Number BC009190, with the NCBI Gene ID 28998 and UniProt ID Q9BYD1 . In normal cells, MRPL13 contributes to maintaining mitochondrial function and cellular homeostasis through its involvement in the mitochondrial translation machinery.

What applications are MRPL13 antibodies suitable for in research?

MRPL13 antibodies have been validated for multiple research applications, making them versatile tools for investigating this protein's expression and function. According to product information and published literature, MRPL13 antibodies are suitable for:

• Western Blot (WB): For detecting MRPL13 protein expression levels in cell and tissue lysates

• Immunoprecipitation (IP): For isolating MRPL13 protein complexes from biological samples

• Immunohistochemistry (IHC): For visualizing MRPL13 expression patterns in tissue sections

• ELISA: For quantitative detection of MRPL13 in various sample types

Published applications have demonstrated successful use of MRPL13 antibodies in Western blot analyses across multiple studies, with at least six publications cited for WB applications and one publication for IHC applications .

Positive Western blot detection has been confirmed in various cell lines and tissues, including HL-60 cells, HeLa cells, Jurkat cells, COS-7 cells, mouse lung tissue, and rat lung tissue . Positive immunoprecipitation has been demonstrated in mouse lung tissue, while positive immunohistochemistry has been shown in human liver cancer tissue and human breast cancer tissue .

What are the optimal dilutions for different experimental applications of MRPL13 antibodies?

The optimal dilutions for MRPL13 antibodies vary depending on the specific application and the antibody being used. Based on product information from manufacturers, the following dilution ranges are recommended:

It is important to note that these are general recommendations, and the optimal dilution may need to be determined empirically for each experimental system. As stated in the product information: "It is recommended that this reagent should be titrated in each testing system to obtain optimal results" .

For immunohistochemistry applications, specific antigen retrieval methods have been suggested, including "antigen retrieval with TE buffer pH 9.0" or alternatively "antigen retrieval may be performed with citrate buffer pH 6.0" . The optimal dilution can be influenced by factors such as the specific antibody preparation, the abundance of MRPL13 in the sample, the detection method being used, and the sample preparation method.

What species reactivity has been confirmed for MRPL13 antibodies?

MRPL13 antibodies have been tested and confirmed to react with MRPL13 proteins from multiple species, making them valuable tools for comparative studies across different model organisms. According to the product information, the tested reactivity includes:

• Human

• Mouse

• Rat

• Monkey

This multi-species reactivity is particularly useful for researchers working with various animal models in their studies of MRPL13 function. The cited reactivity in published literature has primarily focused on human samples, which reflects the significant interest in MRPL13's role in human diseases, particularly cancer .

The cross-reactivity across species suggests a high degree of conservation in the MRPL13 protein structure among mammals, which is consistent with its fundamental role in mitochondrial function. This conservation makes MRPL13 antibodies versatile reagents for comparative studies between human disease models and animal models.

What is the recommended storage condition for MRPL13 antibodies?

For optimal maintenance of antibody activity and stability, MRPL13 antibodies should be stored according to manufacturer recommendations. The consensus storage conditions from the product information include:

• Store at -20°C

• The antibody is stable for one year after shipment when stored properly

• For 20μl sizes containing 0.1% BSA, aliquoting is unnecessary for -20°C storage

• For other preparations, it is advisable to aliquot the antibody to avoid repeated freeze/thaw cycles

• The antibody is typically supplied in a liquid form in PBS buffer with 0.02% sodium azide and 50% glycerol at pH 7.3

Proper storage is critical for maintaining antibody performance over time. Repeated freeze/thaw cycles can lead to degradation of the antibody and reduction in its specificity and sensitivity. Therefore, if the antibody will be used multiple times, it is recommended to prepare small aliquots upon receipt.

What is the molecular weight of MRPL13 protein for Western blot detection?

When performing Western blot analysis for MRPL13, researchers should expect to observe bands corresponding to the molecular weight of the protein. According to the product information:

• The calculated molecular weight of MRPL13 is 21 kDa based on its 178 amino acid sequence

• The observed molecular weight in experimental systems typically ranges from 20-23 kDa

This slight variation between calculated and observed molecular weights is common in protein analysis and may be due to post-translational modifications, protein conformation, or differences in gel electrophoresis conditions. When validating MRPL13 antibodies in Western blot applications, the presence of bands within this 20-23 kDa range would be consistent with specific detection of MRPL13.

How can MRPL13 expression be accurately quantified in cancer tissue compared to normal tissue?

Accurate quantification of MRPL13 expression differences between cancer and normal tissues requires robust methodological approaches. Based on research findings, several complementary techniques can be employed:

• Quantitative PCR (qPCR): For RNA-level analysis, researchers have successfully used qPCR to demonstrate significant differences in MRPL13 expression between cancer tissues (particularly lung adenocarcinoma) and control samples . This approach requires careful RNA extraction, use of appropriate reference genes, specific primer design, and controls to ensure specificity.

• Western Blot Analysis: For protein-level quantification, Western blotting with MRPL13 antibodies provides a semi-quantitative assessment of expression differences. Use 1:500-1:2000 dilution of MRPL13 antibody with appropriate loading controls and densitometry software for quantitative analysis .

• Immunohistochemistry (IHC): For spatial analysis of expression within tissue architecture, use 1:50-1:500 dilution ranges with standardized scoring systems and appropriate controls . Antigen retrieval with TE buffer pH 9.0 or citrate buffer pH 6.0 is recommended.

• Multi-omics Integration: Research has demonstrated that integrating data from multiple platforms provides more robust assessment by correlating RNA-seq data with protein expression and examining methylation status in relation to expression .

Recent studies have shown that MRPL13 expression is significantly higher in various cancer tissues compared to normal tissues at both RNA and protein levels, making it a potential biomarker for cancer diagnosis and prognosis .

What is the relationship between MRPL13 expression and cancer prognosis?

Recent studies have revealed MRPL13 as a significant prognostic indicator across multiple cancer types. Research findings indicate:

• Prognostic Association: Higher MRPL13 expression levels correlate with poorer prognosis in several cancer types, with research concluding that MRPL13 serves as "a molecular biomarker for diagnosing and suggesting the prognosis of certain malignant tumors" .

• Cancer-Specific Patterns: While MRPL13 has pan-cancer implications, its prognostic value has been particularly well-established in lung adenocarcinoma (LUAD) .

• Multivariate Significance: MRPL13 expression maintains independent prognostic value even when controlling for traditional clinical parameters, suggesting its utility in enhancing existing prognostic frameworks.

• Molecular Correlations: MRPL13 expression correlates with specific molecular features with prognostic implications, including methylation characteristics, tumor mutation burden (TMB), microsatellite instability (MSI), and homologous recombination deficiency (HRD) .

• Cellular Process Associations: Single-cell analysis has revealed that MRPL13 expression positively correlates with aggressive cancer phenotypes including metastasis, epithelial-mesenchymal transition (EMT), cell cycle progression, DNA repair, invasion, and proliferation .

The consistent association between elevated MRPL13 expression and poorer outcomes across cancer types highlights its potential as both a prognostic biomarker and therapeutic target . These findings suggest that incorporating MRPL13 expression analysis into clinical assessments could improve risk stratification and treatment planning.

How does MRPL13 knockdown affect cancer cell phenotypes and what methods are most effective?

Research has demonstrated that MRPL13 knockdown produces significant phenotypic changes in cancer cells, particularly in lung adenocarcinoma models. The following methodological approaches and observed effects have been reported:

Effective Knockdown Methods:

• siRNA Transfection: Transient knockdown using small interfering RNA targeting MRPL13, useful for short-term phenotypic assays.

• shRNA Stable Expression: Lentiviral or retroviral delivery of short hairpin RNA targeting MRPL13, enabling selection of stable knockdown cell lines for long-term studies.

• CRISPR/Cas9 Gene Editing: Provides complete knockout rather than knockdown, enabling study of complete loss of MRPL13 function.

Observed Phenotypic Effects:

Research has shown that knockdown of MRPL13 in cancer cells leads to several significant changes:

These findings suggest that MRPL13 may be an important target for anti-cancer therapeutic strategies, particularly in lung adenocarcinoma. When designing MRPL13 knockdown experiments, researchers should carefully consider the duration of study and specific phenotypic endpoints of interest.

What is the relationship between MRPL13 expression and immune infiltration in tumor microenvironments?

The relationship between MRPL13 expression and immune infiltration in the tumor microenvironment represents an important aspect of cancer biology with potential therapeutic implications. Research findings have revealed several significant associations:

• Macrophage Polarization: MRPL13 appears to influence macrophage polarization within the tumor microenvironment, with high MRPL13 expression associated with promotion of macrophage transformation to the M1 state .

• T Cell Infiltration: MRPL13 expression levels inversely correlate with the infiltration of antitumor T cells into the tumor microenvironment. Knockdown of MRPL13 appears to reduce the inhibition of T cell infiltration .

• Correlation with Immune Markers: Pan-cancer analysis has revealed that MRPL13 expression correlates with specific immune infiltration patterns, though the relationship varies across different cancer types .

• Methodological Approaches for Investigation: Various methods can be employed to study this relationship, including bioinformatic analysis of TCGA data, flow cytometry, multiplex immunohistochemistry, and single-cell RNA sequencing.

• Translational Significance: The correlation between MRPL13 and immune infiltration patterns may have implications for immunotherapy response prediction, potentially helping identify patients who might benefit from combined therapies targeting both MRPL13 and immune pathways.

The mechanistic basis for these relationships remains an active area of investigation. Given MRPL13's role in mitochondrial function, it's possible that its effects on energy metabolism in cancer cells indirectly influence the tumor microenvironment and immune cell function.

How can researchers validate the specificity of MRPL13 antibodies in their experimental systems?

Validating antibody specificity is crucial for ensuring reliable and reproducible research results. For MRPL13 antibodies, several complementary validation approaches are recommended:

• Positive and Negative Control Samples:

  • Use samples known to express MRPL13, such as HeLa cells, HL-60 cells, Jurkat cells, COS-7 cells, mouse lung tissue, or rat lung tissue

  • Include samples with known low or no expression of MRPL13

  • The most rigorous approach involves comparing antibody signals between wild-type samples and those where MRPL13 has been depleted through siRNA, shRNA, or CRISPR methods

• Western Blot Band Size Verification:

  • Confirm that the detected band appears at the expected molecular weight (20-23 kDa)

  • Look for a single, clean band rather than multiple bands that might indicate cross-reactivity

• Multiple Antibody Validation:

  • Use multiple antibodies targeting different epitopes of MRPL13

  • Compare staining patterns and ensure consistent results

• Subcellular Localization Consistency:

  • Since MRPL13 is a mitochondrial protein, confirm appropriate mitochondrial localization in immunofluorescence or IHC studies

  • Co-staining with established mitochondrial markers can verify proper localization

• Correlation Between Protein and mRNA Levels:

  • Compare protein detection by the antibody with mRNA expression data

  • While not always perfectly correlated, consistent patterns support antibody specificity

How can single-cell analysis be leveraged to study MRPL13 expression in heterogeneous tumor samples?

Single-cell analysis offers powerful approaches for dissecting MRPL13 expression patterns and functional relationships within the complex heterogeneity of tumor samples. Several methodological considerations are important for researchers planning such studies:

• Single-Cell RNA Sequencing (scRNA-seq) Approaches:

  • Consider platform selection based on desired cell numbers and sequencing depth

  • Optimize tissue dissociation protocols to maintain cell viability

  • Ensure sufficient sequencing depth for reliable detection of MRPL13 transcripts

  • Implement computational workflows that accurately quantify MRPL13 expression at the single-cell level

• Functional Module Correlation Analysis:

  • Research has demonstrated that single-cell data analysis can reveal relationships between MRPL13 expression and functional modules in lung adenocarcinoma

  • Positive correlations have been observed with modules related to metastasis, EMT, cell cycle progression, DNA repair, invasion, DNA damage, and proliferation

  • Negative correlations have been observed with hypoxia and inflammation modules

• Cell Type-Specific Expression Patterns:

  • Apply robust cell typing algorithms to identify major cell populations

  • Assess whether MRPL13 is differentially expressed across distinct cell types within the tumor microenvironment

  • Identify potential cancer cell subpopulations with varying MRPL13 expression

• Spatial Context Integration:

  • Combine single-cell resolution with spatial information using platforms like spatial transcriptomics

  • Use MRPL13 antibodies in spatial protein profiling approaches such as multiplex immunohistochemistry

  • Develop computational methods to integrate spatial and single-cell data

By leveraging these advanced single-cell approaches, researchers can gain unprecedented insights into the heterogeneous expression and function of MRPL13 within the complex tumor microenvironment, potentially revealing new therapeutic opportunities.

What are the methodological considerations for studying MRPL13 in correlation with cuproptosis-related genes?

Recent research has identified important correlations between MRPL13 and cuproptosis-related genes, suggesting a potential link between mitochondrial ribosomal function and this copper-dependent cell death pathway . When investigating these relationships, researchers should consider the following methodological approaches:

• Study Design Considerations:

  • Pan-cancer analysis is valuable as research has indicated that "MRPL13 in pan-cancer is related to cuproptosis-related genes"

  • Consider whether relationships between MRPL13 and cuproptosis genes vary by tissue type or cancer subtype

  • Choose cell lines with varying levels of MRPL13 expression and copper sensitivity

• Expression Correlation Methods:

  • Perform correlation analyses between MRPL13 and established cuproptosis genes using transcriptomic data

  • Use large datasets such as TCGA and GTEx for statistical power

  • Validate findings at the protein level using MRPL13 antibodies in Western blot analysis alongside antibodies for cuproptosis-related proteins

• Functional Relationship Investigation:

  • Knockdown/overexpression of MRPL13 followed by assessment of cuproptosis gene expression

  • Protocols for inducing cuproptosis using copper ionophores followed by assessment of MRPL13 expression changes

• Mitochondrial Function Assessment:

  • Measure oxygen consumption rate (OCR) in cells with varied MRPL13 expression

  • Assess effects of copper treatment on mitochondrial function

  • Evaluate mitochondrial membrane potential changes associated with MRPL13 expression and copper treatment

• Cell Death Assays in Context of MRPL13 Modulation:

  • Assess cuproptosis-specific death markers such as lipid peroxidation and protein lipoylation

  • Distinguish cuproptosis from apoptosis, ferroptosis, and other death pathways

By employing these methodological considerations, researchers can effectively investigate the emerging relationship between MRPL13 and cuproptosis-related pathways, potentially uncovering new therapeutic vulnerabilities in cancers with aberrant MRPL13 expression.