TEX47 Antibody

What is TEX47 and why is it important in research?

TEX47, also known as dynein-associated BLUF protein or uncharacterized protein C7orf62, is a 253 amino acid protein with a molecular mass of approximately 29.5 kDa that shows testis-specific expression in humans . Its importance in research stems from its putative role as a human orthologue of LAX28, a protein required for the stable assembly of the inner dynein arm f complex in flagella . This connection suggests TEX47 may play a critical role in human sperm motility and potentially in other ciliated cells. Understanding TEX47 function could provide insights into certain forms of male infertility and ciliopathies.

What applications are TEX47 antibodies suitable for?

Based on current commercial offerings, TEX47 antibodies are validated for multiple research applications including Western Blotting (WB), immunohistochemistry on frozen sections (IHC-fr), immunohistochemistry on paraffin-embedded tissues (IHC-p), and immunofluorescence (IF) . The availability of differently conjugated antibodies (such as biotin-tagged and Cy5-tagged options) expands the potential applications to include multiplexed detection systems and flow cytometry, though researchers should verify specific validations for their experimental systems.

What tissue expression pattern does TEX47 show?

TEX47 exhibits highly specific expression primarily in testicular tissue . This tissue-restricted pattern makes it particularly valuable for reproductive biology research and as a potential biomarker for testis-specific processes. When designing experiments to study TEX47, researchers should consider using testicular tissue as positive controls and non-reproductive tissues as negative controls to verify antibody specificity.

How can I validate the specificity of a TEX47 antibody?

Antibody validation should include:

• Western blot analysis showing a band at the expected molecular weight (29.5 kDa)

• Comparison of staining between testicular tissue (positive) and non-expressing tissues (negative)

• Knockdown or knockout controls if available

• Peptide competition assays to verify epitope specificity

• Testing multiple antibody clones targeting different epitopes of TEX47

This multi-approach validation strategy helps ensure experimental results reflect true TEX47 biology rather than non-specific binding artifacts.

How does TEX47's structure relate to its function in dynein complexes?

TEX47 shares structural features with sensory BLUF (Blue Light Using FAD) domain-containing proteins , suggesting potential roles in signaling or protein-protein interactions. Research in Leishmania has shown that LAX28 (the TEX47 orthologue) is essential for stable assembly of the inner dynein arm f complex and proper flagellar function . TEX47 likely serves as a structural component that bridges or stabilizes interactions between dynein complex subunits.

Current structural analysis suggests the BLUF domain may function in protein-protein interactions rather than in blue light sensing in this context. Researchers investigating TEX47 function should consider protein-protein interaction studies (co-immunoprecipitation, proximity labeling) to identify binding partners within the dynein complex.

What are the methodological considerations for detecting TEX47 in human sperm samples?

When studying TEX47 in human sperm samples, researchers should consider:

• Sample preparation: Fresh samples should be processed promptly to preserve protein integrity

• Fixation protocols: Optimized paraformaldehyde fixation (typically 4%) is recommended for immunofluorescence studies

• Permeabilization: Due to the compact nature of sperm chromatin, enhanced permeabilization may be necessary (0.5% Triton X-100)

• Antibody selection: Use antibodies validated specifically for sperm immunocytochemistry

• Controls: Include healthy donor samples as positive controls and pre-immune serum as negative controls

For Western blotting applications, specialized sperm protein extraction buffers containing reducing agents and protease inhibitors are essential to overcome the highly crosslinked nature of sperm proteins.

How can TEX47 antibodies be used to investigate ciliopathies?

Given the connection between TEX47 and flagellar/ciliary dynein complexes , TEX47 antibodies can be valuable tools for investigating ciliopathies through:

• Immunohistochemical analysis of ciliated tissues from patients with suspected ciliopathies

• Co-localization studies with other ciliary markers to assess ciliary structure integrity

• Comparative analysis of TEX47 localization in healthy versus diseased ciliated tissues

• Quantitative assessment of TEX47 expression levels in patient samples

• Screening for TEX47 interactions with known ciliopathy-associated proteins

Researchers should employ both tissue and cell culture models of ciliopathies, utilizing TEX47 antibodies to examine potential contributions to disease mechanisms.

What approaches can resolve contradictory results in TEX47 immunodetection studies?

When facing conflicting results in TEX47 immunodetection:

• Compare antibody epitopes: Different antibodies may target distinct regions of TEX47, potentially recognizing different isoforms or post-translationally modified variants

• Evaluate fixation sensitivity: Test multiple fixation protocols as some epitopes may be fixative-sensitive

• Implement orthogonal detection methods: Combine antibody-based detection with mRNA analysis or mass spectrometry

• Consider species differences: If using model organisms, sequence homology and conservation of the epitope region should be verified

• Examine expression conditions: TEX47 expression may be developmentally regulated or influenced by physiological state

A systematic comparison table documenting experimental conditions across studies can help identify sources of variation leading to contradictory results.

What are the optimal conditions for using TEX47 antibodies in Western blotting?

For optimal Western blot detection of TEX47:

• Sample preparation: Use strong lysis buffers (containing SDS and reducing agents) to ensure complete protein extraction

• Gel percentage: 12-15% polyacrylamide gels provide optimal resolution for the 29.5 kDa TEX47 protein

• Transfer conditions: Semi-dry or wet transfer at 100V for 1 hour typically yields efficient transfer

• Blocking: 5% non-fat milk or BSA in TBST for 1 hour at room temperature

• Primary antibody dilution: Typically 1:1000, but optimization is recommended for each antibody

• Incubation: Overnight at 4°C provides the best signal-to-noise ratio

• Detection system: HRP-conjugated secondary antibodies with enhanced chemiluminescence detection offer good sensitivity

These conditions should be optimized for specific experimental systems, as protein extraction efficiency can vary between tissue types.

How can immunofluorescence detection of TEX47 be optimized for co-localization studies?

For optimal co-localization studies with TEX47 antibodies:

• Use TEX47 antibodies with different host species than antibodies for co-localization targets

• For multi-color imaging, select conjugated TEX47 antibodies (such as Cy5-conjugated options ) compatible with your fluorescence filter sets

• Implement spectral unmixing if fluorophore emission spectra overlap

• Employ high-resolution imaging techniques (confocal, structured illumination microscopy) for accurate co-localization assessment

• Calculate co-localization coefficients (Pearson's, Mander's) for quantitative analysis

• Include appropriate controls for non-specific binding and autofluorescence

Researchers should be particularly attentive to fixation protocols, as over-fixation can mask epitopes while under-fixation can result in structural artifacts.

What experimental design is recommended to investigate TEX47's role in flagellar assembly?

A comprehensive experimental design would include:

• Temporal analysis: Examine TEX47 expression and localization during different stages of flagellar development

• Genetic manipulation: CRISPR/Cas9 knockout or knockdown of TEX47 to assess effects on flagellar structure and function

• Rescue experiments: Re-expression of wild-type or mutant TEX47 in knockout models

• Protein-protein interactions: Co-immunoprecipitation or proximity labeling to identify TEX47 binding partners

• Functional assays: Motility analysis in sperm or other flagellated cells with modified TEX47 expression

• Structural studies: Electron microscopy to assess flagellar ultrastructure in the presence/absence of TEX47

This multi-faceted approach would provide comprehensive insights into TEX47's functional role in flagellar dynamics.

How can TEX47 antibodies contribute to male infertility research?

TEX47 antibodies can advance male infertility research through:

• Diagnostic applications: Developing immunoassays to detect TEX47 abnormalities in infertile men

• Phenotypic classification: Categorizing sperm motility defects based on TEX47 expression patterns

• Mechanistic studies: Investigating the molecular basis of TEX47-associated motility dysfunction

• Biomarker development: Evaluating TEX47 as a potential biomarker for specific forms of male infertility

• Therapeutic target assessment: Exploring TEX47-related pathways for potential therapeutic interventions

The testis-specific expression of TEX47 makes it particularly valuable for studies focused on male reproductive biology and pathology.

What approaches can determine if TEX47 has functions beyond flagellar assembly?

To investigate potential additional functions of TEX47:

• Interaction proteomics: Mass spectrometry-based identification of TEX47 binding partners across different tissues

• Subcellular localization studies: High-resolution imaging to determine if TEX47 localizes to structures beyond flagella

• Transcriptomics analysis: RNA-seq following TEX47 manipulation to identify affected pathways

• Evolutionary analysis: Comparative genomics to identify conserved domains that might suggest additional functions

• Non-flagellated cell studies: Examining TEX47 expression and function in cells lacking flagella

Researchers should remain open to unexpected findings, as many proteins initially characterized in specific contexts later reveal broader biological roles.

How can TEX47 antibodies be incorporated into multi-omics research approaches?

TEX47 antibodies can complement multi-omics approaches through:

• Integration with proteomics: Antibody-based enrichment for targeted mass spectrometry

• Validation of transcriptomics findings: Confirming protein-level changes corresponding to TEX47 mRNA alterations

• Chromatin immunoprecipitation (ChIP) studies: If TEX47 has potential nuclear functions

• Tissue-specific interactome mapping: Identifying context-dependent protein interactions

• Spatial transcriptomics validation: Confirming localization of TEX47 expression in complex tissues

This integration enables researchers to connect genomic, transcriptomic, and proteomic data to cellular and tissue-level observations regarding TEX47 biology.

What quality control measures should be implemented when using TEX47 antibodies?

Rigorous quality control for TEX47 antibody use should include:

• Lot-to-lot validation: Comparing performance metrics between antibody lots

• Multiple application testing: Confirming specificity across different experimental contexts (WB, IHC, IF)

• Positive and negative control tissues: Testis tissue (positive) versus non-reproductive tissues (negative)

• Peptide competition: Confirming signal abolishment when antibody is pre-incubated with immunizing peptide

• Knockout validation: Testing antibody in TEX47 knockout systems when available

• Cross-reactivity assessment: Testing against related proteins, particularly other BLUF domain-containing proteins

Researchers should document these validation steps in publications to enhance reproducibility and reliability of findings.

What are the considerations for choosing between different TEX47 antibody conjugates?

When selecting between different TEX47 antibody conjugates (e.g., biotin, Cy5) , researchers should consider:

• Detection system compatibility: Ensure conjugate works with available detection instruments

• Multiplexing requirements: Select conjugates that permit simultaneous detection of multiple targets

• Sensitivity needs: Some conjugates offer enhanced signal amplification capabilities

• Tissue autofluorescence profile: Choose fluorophores with emission spectra distinct from sample autofluorescence

• Stability requirements: Consider photobleaching properties for extended imaging sessions

• Application-specific optimization: Different conjugates may perform optimally in different applications

The choice should be guided by specific experimental requirements and available detection systems.