MRPL11 Antibody

What is MRPL11 and why is it significant in mitochondrial research?

MRPL11 (Mitochondrial Ribosomal Protein L11, also known as uL11m) is a critical 39S subunit component of the mitochondrial ribosome encoded by the nuclear genome. It plays an essential role in oxidative phosphorylation and ensures correct assembly and function of the mitochondrial machinery influencing energy demands through connections to translation regulation and metabolic balance . Studying MRPL11 is particularly important because abnormal expression has been associated with mitochondrial encephalopathy, making it relevant for both basic science and disease research .

What types of MRPL11 antibodies are available for research applications?

Multiple types of MRPL11 antibodies are available for research, with variations in host species, clonality, and applications:

The choice between polyclonal antibodies (offering broader epitope recognition) and monoclonal antibodies (providing specificity) should be based on experimental requirements for sensitivity versus reproducibility .

What is the molecular weight of MRPL11 and how does this affect antibody validation?

The calculated molecular weight of MRPL11 is approximately 20-21 kDa, with the observed molecular weight typically being 21 kDa in Western blot applications . When validating MRPL11 antibodies, researchers should confirm that the detected band appears at the expected molecular weight. Alternative splice variants may result in multiple isoforms, so knowledge of your specific experimental system is essential for proper interpretation of results .

What are the optimal dilution ratios for different MRPL11 antibody applications?

Optimal dilution ratios vary by application technique and specific antibody preparation:

These are general guidelines, and optimization is required for each experimental system. Begin with the manufacturer's recommended dilution and adjust based on signal-to-noise ratio in your specific samples .

What antigen retrieval methods are most effective for MRPL11 immunohistochemistry?

For optimal MRPL11 detection in formalin/PFA-fixed paraffin-embedded tissue sections, heat-mediated antigen retrieval is recommended. Two buffer systems have proven effective:

• TE buffer pH 9.0 (primary recommendation)

• Citrate buffer pH 6.0 (alternative option)

Heat-mediated antigen retrieval helps expose epitopes masked during fixation. For human liver tissue samples, a dilution of 1:20-1:200 of the primary antibody following antigen retrieval has shown positive IHC results .

How should MRPL11 antibodies be stored to maintain activity?

To maintain optimal activity of MRPL11 antibodies:

• Store at -20°C in aliquots to minimize freeze-thaw cycles

• Most preparations are stable for one year after shipment when properly stored

• For antibodies in liquid form, storage buffers typically include:

  • PBS with 0.02% sodium azide and 50% glycerol (pH 7.3)

  • Some formulations may contain BSA (0.1%)

Proper storage is critical as repeated freeze-thaw cycles can significantly reduce antibody activity and specificity .

How can MRPL11 antibodies be utilized in mitochondrial disease research?

MRPL11 antibodies serve as valuable tools in investigating mitochondrial disorders, particularly mitochondrial encephalopathy where MRPL11 expression is downregulated . Research applications include:

• Comparative expression analysis of MRPL11 between healthy and diseased tissues

• Immunoprecipitation combined with mass spectrometry to identify MRPL11 interaction partners

• Immunohistochemistry to localize MRPL11 in tissue sections from patients with mitochondrial disorders

• Co-immunoprecipitation to investigate protein-protein interactions involved in mitochondrial ribosome assembly

These approaches help elucidate the molecular mechanisms underlying mitochondrial diseases and potential therapeutic targets .

What controls should be included when using MRPL11 antibodies in multiplexed immunoassays?

For reliable results in multiplexed immunoassays using MRPL11 antibodies:

• Positive controls: Include known MRPL11-expressing cell lines such as HepG2, HeLa, K-562, or BxPC-3 cells

• Negative controls:

  • Isotype control (rabbit monoclonal IgG or mouse IgG2a depending on antibody host)

  • Unlabeled control (cells without primary and secondary antibody incubation)

• Technical controls:

  • Signal specificity validation using MRPL11 knockdown or knockout samples

  • Cross-reactivity assessment when using multiple antibodies simultaneously

These controls are essential for distinguishing true positive signals from background and non-specific binding, particularly in complex multiplexed systems .

How do different cell fixation methods affect MRPL11 antibody performance in immunofluorescence?

Cell fixation methodology significantly impacts MRPL11 antibody performance in immunofluorescence applications:

• Paraformaldehyde fixation (4%):

  • Recommended for preserving cellular architecture

  • Requires 10-minute fixation followed by permeabilization with 0.1% PBS-Tween

  • Works well with most MRPL11 antibodies at 1:50-1:500 dilution

• Methanol fixation (80-90%):

  • Alternative approach that simultaneously fixes and permeabilizes

  • 5-minute fixation is typically sufficient

  • Particularly effective for intracellular flow cytometry applications

The choice between these methods should be based on the specific cellular compartment being studied and the particular epitope recognized by your antibody .

What are common causes of non-specific binding when using MRPL11 antibodies?

When encountering non-specific binding with MRPL11 antibodies, consider these common causes and solutions:

• Antibody concentration too high: Optimize dilution; try increasing from 1:1000 to 1:2000 or higher

• Insufficient blocking: Extend blocking time using 10% normal serum with 0.3M glycine

• Cross-reactivity with similar proteins: Verify antibody specificity using knockout/knockdown controls

• Sample preparation issues: Ensure complete lysis and denaturation for Western blots

• Buffer compatibility problems: Some antibodies perform better in specific buffer systems

Methodical optimization of each experimental parameter and inclusion of appropriate controls can significantly reduce non-specific binding .

How can researchers distinguish between MRPL11 isoforms resulting from alternative splicing?

To differentiate between MRPL11 isoforms resulting from alternative splicing:

• Western blotting with high-resolution gels: Use 12-15% polyacrylamide gels to resolve small molecular weight differences

• Isoform-specific antibodies: Select antibodies raised against unique regions of specific isoforms

• RT-PCR analysis: Complement protein detection with transcript analysis using isoform-specific primers

• Mass spectrometry: For definitive identification of protein isoforms

• Bioinformatic analysis: Cross-reference observed molecular weights with predicted weights of known isoforms

This multi-faceted approach provides comprehensive characterization of MRPL11 isoform expression patterns in your experimental system .

What strategies can resolve discrepancies in MRPL11 detection between different antibodies?

When facing inconsistent results between different MRPL11 antibodies:

• Epitope mapping: Determine the specific regions recognized by each antibody

• Validation using multiple techniques: Compare results across WB, IHC, and IF applications

• Cell type considerations: Some antibodies perform better in certain cell types (e.g., HepG2 vs. HeLa)

• Protocol standardization: Ensure identical sample preparation, blocking, and detection methods

• Post-translational modifications: Consider whether PTMs might mask epitopes in certain contexts

Systematic comparison using standardized positive controls (e.g., recombinant MRPL11) can help identify the most reliable antibody for your specific application .

How does MRPL11 antibody staining correlate with mitochondrial function assays?

Correlation between MRPL11 antibody staining and functional mitochondrial assays provides valuable mechanistic insights:

• Oxygen consumption rate (OCR): MRPL11 expression levels often correlate with mitochondrial respiratory capacity

• ATP production: Changes in MRPL11 levels may reflect alterations in oxidative phosphorylation efficiency

• Mitochondrial membrane potential: Combined MRPL11 immunofluorescence with membrane potential dyes (e.g., TMRM) can reveal relationships between ribosomal integrity and mitochondrial function

• mtDNA-encoded protein synthesis: MRPL11 staining intensity may predict translation efficiency of mitochondrially-encoded proteins

This integrated approach provides a comprehensive assessment of how MRPL11 abundance affects mitochondrial functional outputs in various research contexts .

What emerging applications utilize MRPL11 antibodies in studying mitochondrial quality control mechanisms?

Cutting-edge applications of MRPL11 antibodies in mitochondrial quality control research include:

• Mitophagy assessment: Dual labeling with MRPL11 and autophagy markers to track mitochondrial degradation

• Stress response monitoring: Evaluating MRPL11 expression changes during mitochondrial unfolded protein response

• Dynamic mitochondrial remodeling: Time-lapse imaging with MRPL11 antibodies to observe mitochondrial network changes

• Single-cell analysis: Flow cytometry applications to correlate MRPL11 levels with mitochondrial mass and function at the single-cell level

• Proximity labeling approaches: Using MRPL11 antibodies in conjunction with techniques like BioID to map the spatial organization of mitochondrial ribosomes

These approaches provide new insights into fundamental mitochondrial biology and disease mechanisms .

How do MRPL11 expression patterns differ across tissue types and disease states?

Research using MRPL11 antibodies has revealed tissue-specific and disease-related expression patterns:

• Tissue specificity:

  • High expression in metabolically active tissues (liver, kidney, heart)

  • Variable expression in neuronal tissues

  • Detectable in cultured cell lines including HepG2, HeLa, K-562, and BxPC-3

• Disease associations:

  • Downregulation in mitochondrial encephalopathy

  • Altered expression in various cancers

  • Potential biomarker for mitochondrial dysfunction in neurodegenerative conditions