TRIM41 Antibody

What is TRIM41 and what are its primary functions?

TRIM41 is an E3 ubiquitin ligase belonging to the TRIM (tripartite motif) family of proteins. Its primary function involves targeting specific proteins for ubiquitination and subsequent degradation via the proteasome pathway. Research has identified two significant roles for TRIM41:

• Immune regulation: TRIM41 can induce c-Maf degradation and interfere with IL-10 expression in airway dendritic cells (DCs), compromising their tolerogenic functions in airway allergy models .

• Intrinsic immunity: TRIM41 acts as an intrinsic immune factor against vesicular stomatitis virus (VSV) by targeting the viral nucleoprotein for ubiquitination and subsequent proteasomal degradation .

Understanding these functions provides context for the utility of TRIM41 antibodies in investigating immune regulation and antiviral defense mechanisms.

What applications are TRIM41 antibodies suitable for?

Based on available research, TRIM41 antibodies have been validated for multiple applications:

Researchers should note that validation is typically performed using cell lysates as positive controls, and application-specific optimization may be required for different experimental systems .

What species reactivity can be expected with TRIM41 antibodies?

Commercial TRIM41 antibodies show cross-reactivity with multiple species. According to the provided information, antibodies targeting different epitopes of TRIM41 have demonstrated reactivity with:

• Human (100% predicted reactivity)

• Mouse (100% predicted reactivity)

• Rat (100% predicted reactivity)

• Dog (93% predicted reactivity)

• Horse (100% predicted reactivity)

• Cow (86% predicted reactivity)

• Rabbit (93% predicted reactivity)

• Pig (100% predicted reactivity)

• Zebrafish (90% predicted reactivity)

• Yeast (Saccharomyces cerevisiae) (90% predicted reactivity)

This broad cross-reactivity makes TRIM41 antibodies valuable tools for comparative studies across species.

How does TRIM41 contribute to the pathogenesis of airway allergy, and how can antibodies help investigate this mechanism?

TRIM41 plays a significant role in airway allergy pathogenesis through multiple mechanisms:

• Increased TRIM41 expression in airway dendritic cells (DCs) of airway allergy (AA) mice correlates with compromised DC tolerogenic functions .

• TRIM41 induces c-Maf degradation, which interferes with IL-10 expression in airway DCs. This mechanism has been demonstrated through immunoprecipitation with anti-TRIM41 antibodies revealing a complex of TRIM41 and c-Maf .

• The expression of TRIM41 in airway DCs positively correlates with airway allergy responses, including increased eosinophil peroxidase, mast cell protease-1, Th2 cytokines, and specific IgE in bronchoalveolar lavage fluids .

Researchers can use TRIM41 antibodies to:

• Quantify TRIM41 expression levels in airway DCs via Western blotting

• Identify protein-protein interactions involving TRIM41 using co-immunoprecipitation

• Visualize TRIM41 localization in airway tissues using immunohistochemistry

• Track changes in TRIM41 expression following experimental interventions

Understanding these mechanisms may lead to novel therapeutic approaches, as inhibition of TRIM41 has been shown to mitigate experimental airway allergy responses .

What is the role of TRIM41 in antiviral immunity, and how can antibodies be used to investigate this function?

TRIM41 has been identified as an intrinsic immune factor against vesicular stomatitis virus (VSV). The mechanism of this antiviral activity has been characterized as follows:

• TRIM41 interacts with the nucleoprotein of VSV (VSV-N), as demonstrated through co-immunoprecipitation studies .

• This interaction leads to ubiquitination of VSV-N, which has been demonstrated both in cellulo and in vitro .

• The ubiquitination of VSV-N by TRIM41 targets it for proteasomal degradation, thereby limiting VSV replication .

Both gain-of-function and loss-of-function studies have confirmed TRIM41's role in restricting VSV infection. Overexpression of TRIM41 inhibits VSV replication activity and reduces viral titers, while knockdown of TRIM41 enhances VSV infection .

Researchers can use TRIM41 antibodies to:

• Evaluate endogenous TRIM41 expression levels in different cell types

• Investigate the interaction between TRIM41 and viral proteins

• Monitor changes in TRIM41 expression during viral infection

• Assess the correlation between TRIM41 levels and viral resistance

How can TRIM41 antibodies be used to differentiate between TRIM41 isoforms or post-translational modifications?

TRIM41 antibodies targeting different epitopes can be valuable tools for investigating various aspects of TRIM41 biology:

• N-terminal vs. C-terminal antibodies: Commercial antibodies targeting different regions of TRIM41 are available, including N-terminal (e.g., AA sequences CGHNFCRVCV TQLWGGEDEE DRDELDREEE EEDGEEEEVE AVGAGAGWDT) and C-terminal epitopes . These can be used to:

  • Detect specific isoforms that may lack certain domains

  • Investigate protein processing or cleavage products

  • Confirm full-length protein expression

• Post-translational modifications: Since TRIM41 functions as an E3 ubiquitin ligase, it may itself be subject to regulation via post-translational modifications. Researchers can:

  • Use phospho-specific antibodies (if available) to detect activation states

  • Employ co-immunoprecipitation with anti-TRIM41 antibodies followed by detection with anti-ubiquitin antibodies to assess TRIM41 auto-ubiquitination

  • Combine TRIM41 antibodies with mass spectrometry approaches to identify novel modifications

This approach enables more detailed investigation of TRIM41 regulation and function in different cellular contexts.

What are the optimal conditions for using TRIM41 antibodies in Western blotting experiments?

For optimal Western blotting results with TRIM41 antibodies, researchers should consider:

• Sample preparation:

  • Use appropriate lysis buffers containing protease inhibitors to prevent TRIM41 degradation

  • Consider adding deubiquitinase inhibitors when investigating TRIM41-mediated ubiquitination

  • Include phosphatase inhibitors if investigating phosphorylation-dependent functions

• Gel selection and transfer:

  • TRIM41 has a molecular weight of approximately 70 kDa

  • Use 8-10% polyacrylamide gels for optimal resolution

  • Transfer to PVDF membranes for stronger protein binding and signal

• Blocking and antibody incubation:

  • Use 5% non-fat dry milk or BSA in TBST for blocking

  • Optimize primary antibody dilution (typically 1:1000 to 1:5000)

  • Incubate primary antibody overnight at 4°C for best results

  • Use HRP-conjugated secondary antibodies at 1:5000 to 1:10000 dilution

• Controls:

  • Include positive controls such as cell lysates known to express TRIM41

  • Consider TRIM41 knockout or knockdown samples as negative controls

  • Use recombinant TRIM41 protein as a size reference when available

These conditions should be optimized for each specific TRIM41 antibody and experimental system.

How can TRIM41 antibodies be used to investigate protein-protein interactions in immune regulation?

TRIM41 antibodies are valuable tools for investigating protein-protein interactions, particularly in the context of immune regulation:

• Co-immunoprecipitation (Co-IP):

  • Use anti-TRIM41 antibodies to precipitate TRIM41 and its interacting partners

  • As demonstrated in research, this approach successfully identified the interaction between TRIM41 and c-Maf in dendritic cells

  • The reverse approach using antibodies against suspected interacting partners (e.g., c-Maf) can confirm the interaction

• Proximity ligation assay (PLA):

  • Combines TRIM41 antibodies with antibodies against potential interaction partners

  • Provides spatial information about interactions within cells

  • Can detect transient or weak interactions that might be lost during Co-IP

• Immunofluorescence co-localization:

  • Use fluorescently labeled TRIM41 antibodies in combination with antibodies against potential interacting proteins

  • Assess co-localization through confocal microscopy

  • Research has demonstrated co-localization of ubiquitin and K48 with c-Maf in the presence of TRIM41

• Pull-down assays with recombinant proteins:

  • Verify direct interactions identified through antibody-based methods

  • Can be combined with in vitro ubiquitination assays to assess functional interactions

These approaches have successfully demonstrated TRIM41's interaction with c-Maf in the context of IL-10 regulation in dendritic cells, as well as its interaction with viral nucleoproteins in antiviral immunity .

What are common challenges when using TRIM41 antibodies and how can they be addressed?

Researchers may encounter several challenges when working with TRIM41 antibodies:

• Specificity issues:

  • Challenge: Cross-reactivity with other TRIM family members due to conserved domains

  • Solution: Validate antibody specificity using TRIM41 knockout/knockdown samples

  • Alternative: Use multiple antibodies targeting different epitopes of TRIM41 to confirm results

• Detection sensitivity:

  • Challenge: Low endogenous expression levels in certain cell types

  • Solution: Consider enrichment methods such as immunoprecipitation before detection

  • Alternative: Use more sensitive detection systems such as chemiluminescent substrates with longer exposure times

• Epitope masking:

  • Challenge: Protein-protein interactions or post-translational modifications may block antibody binding sites

  • Solution: Test multiple antibodies targeting different regions of TRIM41

  • Alternative: Consider denaturing conditions for Western blotting applications

• Degradation during sample preparation:

  • Challenge: TRIM41, as an E3 ligase, may be subject to autoubiquitination and rapid turnover

  • Solution: Include proteasome inhibitors (e.g., MG132) in lysis buffers

  • Alternative: Use shorter sample processing times and maintain samples at 4°C

Detailed troubleshooting requires consideration of the specific application and experimental context.

How can researchers validate TRIM41 antibody specificity for their experimental systems?

Validating antibody specificity is crucial for obtaining reliable results. For TRIM41 antibodies, the following validation approaches are recommended:

• Genetic validation:

  • Use TRIM41 knockout cells or tissues (as described in the VSV study)

  • Employ TRIM41 knockdown using siRNA or shRNA approaches

  • Compare signal intensity between wild-type and knockout/knockdown samples

• Overexpression validation:

  • Express tagged recombinant TRIM41 (e.g., FLAG-tagged TRIM41)

  • Detect with both anti-TRIM41 and anti-tag antibodies

  • Confirm co-localization of signals in immunofluorescence or concordant bands in Western blotting

• Peptide competition:

  • Pre-incubate TRIM41 antibody with the immunizing peptide

  • Compare staining/detection with and without peptide competition

  • Specific signal should be abolished or significantly reduced with peptide competition

• Cross-validation with multiple antibodies:

  • Use antibodies targeting different epitopes of TRIM41

  • Consistent results with different antibodies increase confidence in specificity

  • Consider antibodies from different host species or different clones

These validation approaches should be documented and reported in publications to enhance reproducibility.

What emerging applications of TRIM41 antibodies might advance our understanding of immune regulation?

Several emerging applications of TRIM41 antibodies could significantly advance our understanding of immune regulation:

• Single-cell analysis:

  • Using TRIM41 antibodies for mass cytometry (CyTOF) to analyze TRIM41 expression at single-cell resolution

  • Combining with other immune markers to identify correlations between TRIM41 levels and immune cell subtypes or activation states

  • Investigating heterogeneity in TRIM41 expression within dendritic cell populations

• In vivo imaging:

  • Developing fluorescently labeled TRIM41 antibodies or antibody fragments for in vivo imaging

  • Tracking TRIM41 expression changes during immune responses in real-time

  • Correlating TRIM41 dynamics with disease progression or resolution

• Therapeutic targeting validation:

  • Using antibodies to validate accessibility of TRIM41 for therapeutic targeting

  • Developing antibody-drug conjugates to specifically modulate TRIM41-expressing cells

  • Monitoring changes in TRIM41 expression as a biomarker for treatment response

• Structural studies:

  • Using antibodies that recognize specific conformational states of TRIM41

  • Investigating domain-specific functions through selective blocking antibodies

  • Combining with cryo-EM approaches to understand TRIM41 complex formation

These applications could provide deeper insights into TRIM41's role in immune tolerance and inflammatory diseases beyond the currently established functions in airway allergy and antiviral defense .

How might TRIM41 antibodies be used to explore the intersection of TRIM41's roles in allergic disease and viral immunity?

The dual role of TRIM41 in allergic diseases and antiviral immunity presents an intriguing area for investigation where TRIM41 antibodies could be particularly valuable:

• Temporal dynamics studies:

  • Using TRIM41 antibodies to track expression changes during viral infections in allergic versus non-allergic individuals

  • Investigating whether viral infections alter TRIM41 expression in dendritic cells and affect subsequent allergic responses

  • Examining how allergen exposure affects TRIM41-mediated antiviral functions

• Mechanistic investigations:

  • Determining whether the same pools of TRIM41 are involved in both functions using domain-specific antibodies

  • Investigating whether TRIM41's substrate specificity changes under different immune conditions

  • Using co-immunoprecipitation with TRIM41 antibodies to identify differential interaction partners in allergic versus antiviral responses

• Translational research:

  • Using TRIM41 antibodies to assess expression levels in patient samples

  • Correlating TRIM41 expression with disease severity in allergic patients with or without viral infections

  • Evaluating TRIM41 as a potential biomarker for predicting infection susceptibility in allergic individuals

This research direction could yield insights into the often-observed clinical relationship between viral infections and allergy exacerbations, potentially revealing TRIM41 as a molecular link between these phenomena .