MRPL37 Antibody

What is MRPL37 and why is it important in mitochondrial biology?

MRPL37 has a calculated molecular weight of approximately 48 kDa, which is consistently observed in Western blot applications across multiple antibody vendors . This consistent molecular weight detection is important for experimental validation. When performing Western blot analysis, researchers should expect to observe bands at this molecular weight. Deviation from this expected size may indicate post-translational modifications, proteolytic cleavage, or potential non-specific binding of the antibody .

What are the optimal conditions for using MRPL37 antibodies in Western blot applications?

For optimal Western blot results with MRPL37 antibodies, the following methodological approach is recommended:

• Sample Preparation: Use standard protein extraction protocols with protease inhibitors to prevent degradation of mitochondrial proteins.

• Protein Loading: Load 25-50 μg of total protein per lane for optimal signal detection.

• Dilution Ranges: Different antibodies require different dilutions:

  • Proteintech antibody (15190-1-AP): 1:500-1:1000

  • Proteintech antibody (29522-1-AP): 1:1000-1:3000

  • ABclonal antibody (A4724): 1:200-1:2000

• Blocking Conditions: Use 3-5% nonfat dry milk in TBST for reduced background signal.

• Detection Method: HRP-conjugated secondary antibodies with ECL detection systems are commonly used.

• Exposure Time: Typically 30 seconds to 2 minutes provides optimal signal .

Critically, antibody validation should include positive and negative controls to ensure specificity of binding.

How should researchers optimize MRPL37 antibodies for immunohistochemistry applications?

Optimizing MRPL37 antibodies for immunohistochemistry requires careful consideration of several methodological factors:

• Tissue Processing: Formalin-fixed paraffin-embedded (FFPE) sections are commonly used.

• Antigen Retrieval: Two main methods show effectiveness:

  • TE buffer (pH 9.0) is the primary recommended method for most MRPL37 antibodies

  • Citrate buffer (pH 6.0) can be used as an alternative

• Antibody Dilutions:

  • Abcam antibody (ab224467): 1:20 dilution for FFPE tissues

  • Proteintech antibody (29522-1-AP): 1:150-1:600

  • Sigma-Aldrich antibody (HPA025767): 1:20-1:50

• Signal Detection: Standard DAB (3,3′-diaminobenzidine) detection systems are compatible.

• Expected Staining Pattern: MRPL37 typically shows granular cytoplasmic positivity consistent with mitochondrial localization .

Immunohistochemical staining of human tissues reveals strong granular cytoplasmic positivity in glandular cells of duodenum, which serves as a positive control tissue .

What are the critical considerations for immunofluorescence detection of MRPL37?

For successful immunofluorescence detection of MRPL37, researchers should consider:

• Cell Fixation: PFA-fixation (4% paraformaldehyde) followed by Triton X-100 permeabilization has been validated for successful staining .

• Antibody Dilutions:

  • Abcam antibody (ab224467): 4 μg/ml

  • Proteintech antibody (29522-1-AP): 1:200-1:800

• Counterstains: DAPI nuclear counterstain helps visualize subcellular localization relative to nuclei.

• Expected Pattern: Punctate cytoplasmic staining consistent with mitochondrial localization should be observed.

• Cell Types: U-2 OS (osteosarcoma) and HepG2 cells have been validated for immunofluorescence detection of MRPL37 .

Mitochondrial co-localization markers can be used to confirm the specificity of MRPL37 staining patterns.

How do post-translational modifications of MRPL37 impact antibody selection and experimental design?

MRPL37 undergoes multiple post-translational modifications that can significantly impact antibody recognition and experimental outcomes. According to Uniprot data, the following PTMs have been identified :

When selecting antibodies for PTM-specific research:

• Consider epitope location relative to known PTM sites

• Use phosphatase/deacetylase inhibitors in lysis buffers to preserve PTMs

• Consider PTM-specific antibodies for studies focusing on MRPL37 regulation

• Be aware that heavy ubiquitination may alter migration patterns in Western blots

Antibodies targeting regions containing these modifications may show differential binding depending on the PTM status, potentially leading to varied results across experimental conditions or tissue types .

What are the potential cross-reactivity concerns with MRPL37 antibodies and how can they be addressed?

Cross-reactivity is a significant concern with MRPL37 antibodies due to sequence homology with other mitochondrial ribosomal proteins. To address this:

• Sequence Analysis: MRPL37 shares some sequence similarity with other mitochondrial ribosomal proteins. Detailed BLAST analysis of the immunogen sequence can predict potential cross-reactivity.

• Validation Controls:

  • Use siRNA knockdown or CRISPR knockout of MRPL37 to confirm antibody specificity

  • Include tissues or cell lines known to express varied levels of MRPL37

  • Compare staining patterns with multiple antibodies targeting different epitopes

• Preabsorption Controls: For critical experiments, preabsorption with the immunizing peptide can confirm binding specificity.

• Species Considerations: While many MRPL37 antibodies show cross-reactivity between human, mouse, and rat, the sequence homology is not complete. When working across species, validation experiments should be performed in each species .

How should researchers interpret differences in MRPL37 antibody staining patterns between normal and disease tissues?

Interpretation of differential MRPL37 staining requires careful consideration of multiple factors:

• Mitochondrial Content: Variations may reflect differences in mitochondrial content rather than specific MRPL37 regulation.

• Tissue-Specific Expression: The Human Protein Atlas data shows variable expression across tissues. For example, strong granular cytoplasmic positivity is observed in glandular cells of duodenum .

• Disease Context Analysis:

  • In cancer tissues, altered MRPL37 staining may reflect metabolic reprogramming

  • Changes in staining intensity must be quantified using appropriate controls

  • Compare staining patterns with other mitochondrial markers to distinguish MRPL37-specific changes from general mitochondrial alterations

• Technical Considerations:

  • Antigen retrieval efficiency may vary between normal and disease tissues

  • Fixation artifacts can impact epitope accessibility

  • Background staining should be carefully assessed in each tissue type

Quantitative analysis methods (e.g., digital pathology tools) should be employed for objective comparison between normal and disease samples .

What cell and tissue types are most suitable for studying MRPL37 localization and function?

Based on validation data from multiple antibody suppliers, the following models provide robust systems for MRPL37 research:

• Cell Lines:

  • Human: HeLa, HepG2, K-562, U-251, and U-2 OS cells demonstrate consistent MRPL37 expression

  • These cells show the expected mitochondrial localization pattern

• Tissue Types:

  • Human tissues with strong MRPL37 expression include duodenum, rectum, urinary bladder, and fallopian tube

  • Liver, heart, ovary, and uterus tissues from mice show detectable MRPL37 levels

• Disease Models:

  • Human lymphoma tissue and stomach cancer tissue have been validated for MRPL37 detection

  • These pathological samples may provide insights into disease-related alterations

When selecting experimental models, researchers should consider mitochondrial content and metabolic activity of the tissue type in relation to their specific research questions .

How can researchers effectively combine MRPL37 antibodies with other mitochondrial markers for co-localization studies?

For effective co-localization studies involving MRPL37 and other mitochondrial markers:

• Compatible Marker Selection:

  • Outer membrane markers: TOMM20, VDAC

  • Matrix markers: HSP60, MRPS18B (small mitochondrial ribosomal subunit)

  • Functional markers: OXPHOS complex components (ATP5A, UQCRC2)

• Antibody Compatibility:

  • Select primary antibodies from different host species (e.g., rabbit anti-MRPL37 with mouse anti-TOMM20)

  • If using antibodies from the same species, consider directly conjugated antibodies or sequential staining protocols

• Imaging Considerations:

  • Super-resolution microscopy techniques (STED, STORM) provide enhanced resolution of mitochondrial structures

  • Confocal z-stack imaging is essential for accurate co-localization analysis

  • Deconvolution may improve signal-to-noise ratio

• Quantitative Analysis:

  • Use Pearson's or Mander's correlation coefficients to quantify co-localization

  • Analyze multiple cells and fields to ensure representative results

  • Consider mitochondrial dynamics and potential heterogeneity of staining patterns

This approach enables detailed investigation of MRPL37's precise sub-mitochondrial localization and its relationship to other mitochondrial components .

What are the advanced approaches for studying MRPL37's role in mitochondrial translation using antibody-based techniques?

Advanced research into MRPL37's functional role in mitochondrial translation can be approached through several antibody-dependent techniques:

• Ribosome Profiling with Immunoprecipitation:

  • Use MRPL37 antibodies to isolate intact mitochondrial ribosomes

  • Analyze associated mRNAs to identify translation patterns

  • Compare results under different cellular conditions or disease states

• Proximity Labeling Techniques:

  • APEX2 or BioID fusion with MRPL37 followed by antibody detection of biotinylated proteins

  • This approach identifies proteins in close proximity to MRPL37

  • Can reveal dynamic interaction partners during translation

• Translation Elongation Analysis:

  • Combine puromycin labeling with MRPL37 immunofluorescence

  • This technique allows visualization of active translation sites in relation to MRPL37 localization

  • Can be used to study translation impairment in disease models

• Cross-linking Immunoprecipitation (CLIP):

  • UV cross-linking followed by MRPL37 immunoprecipitation

  • Sequence associated RNAs to identify binding preferences

  • Provides insights into mRNA selectivity of mitochondrial ribosomes

These methods allow researchers to move beyond localization studies to understand the functional significance of MRPL37 in mitochondrial translation dynamics .

What new methodologies are emerging for enhanced detection of MRPL37 in complex biological samples?

Recent advancements in MRPL37 detection include:

• Enhanced Validation Technologies:

  • Orthogonal RNAseq validation confirms antibody specificity by correlating staining intensity with mRNA expression levels

  • Independent validation approaches using multiple antibody types strengthen confidence in results

• Multiplex Imaging Platforms:

  • Cyclic immunofluorescence (CyCIF) allows detection of MRPL37 alongside numerous other proteins

  • Mass cytometry-based imaging techniques (IMC, MIBI) enable simultaneous detection of dozens of proteins including MRPL37

  • These approaches provide unprecedented contextual information about MRPL37 in relation to cellular pathways

• Single-Cell Applications:

  • Antibody-based single-cell proteomics techniques are being adapted for mitochondrial protein detection

  • These methods provide insights into cell-to-cell variation in MRPL37 expression and localization

• Automation and Standardization:

  • Automated staining platforms improve reproducibility of MRPL37 detection

  • Machine learning algorithms enhance quantitative analysis of staining patterns

These developments are driving more reliable and information-rich analysis of MRPL37 in diverse biological contexts .

How can researchers integrate MRPL37 antibody data with other -omics approaches for comprehensive mitochondrial studies?

Integration of antibody-based MRPL37 detection with other -omics approaches requires careful methodological consideration:

• Proteomics Integration:

  • Correlate immunostaining intensity with mass spectrometry-based quantification

  • Use fractionation techniques to enrich mitochondrial samples before analysis

  • Compare post-translational modifications identified by antibodies with PTM proteomics data

• Transcriptomics Correlation:

  • Analyze correlation between MRPL37 protein levels (detected by antibodies) and mRNA expression

  • Identify potential post-transcriptional regulation mechanisms

  • Spatial transcriptomics can be correlated with immunohistochemistry data for tissue-level analysis

• Functional Genomics Approach:

  • Combine CRISPR screening data with antibody-based phenotypic assays

  • Correlate genetic perturbations of MRPL37 with changes in mitochondrial function

  • Use antibodies to validate protein-level changes following genetic manipulation

• Computational Integration Methods:

  • Employ machine learning algorithms to identify patterns across multi-omics datasets

  • Network analysis can position MRPL37 within broader mitochondrial functional pathways

  • Visualization tools can represent complex relationships between different data types

This integrated approach provides a systems-level understanding of MRPL37's role in mitochondrial biology and disease mechanisms .

What are the current challenges in standardizing MRPL37 antibody-based assays across research laboratories?

Standardization of MRPL37 antibody assays faces several challenges that require methodological solutions:

• Antibody Variability:

  • Different epitopes targeted by various commercial antibodies

  • Lot-to-lot variation can impact reproducibility

  • Solution: Implement rigorous validation protocols for each new antibody lot

• Protocol Differences:

  • Variations in fixation, antigen retrieval, and detection methods

  • Sample preparation inconsistencies

  • Solution: Develop detailed standard operating procedures with positive control samples

• Quantification Challenges:

  • Subjective interpretation of staining intensity

  • Different image acquisition settings

  • Solution: Use digital pathology tools and automated scoring systems

• Biological Variables:

  • Cell state fluctuations impact mitochondrial proteins

  • Tissue heterogeneity affects interpretation

  • Solution: Include multiple biological replicates and document experimental conditions thoroughly

• Data Reporting Standards:

  • Inconsistent documentation of experimental conditions

  • Limited sharing of validation data

  • Solution: Adopt standardized reporting formats (e.g., ARRIVE guidelines for animal studies)