IFRD2 Antibody

What are the primary experimental applications for IFRD2 antibodies?

IFRD2 antibodies are primarily used in Western blot (WB), immunohistochemistry on paraffin-embedded tissues (IHC-P), and enzyme-linked immunosorbent assay (ELISA) applications. Commercial polyclonal antibodies have been validated for these techniques, particularly with human samples . When performing Western blot analysis, IFRD2 antibodies can detect the predicted band size of approximately 55 kDa in various human cell lines including HeLa and PC-3 . For IHC-P applications, these antibodies have been successfully used to stain human placental tissue at dilutions of approximately 1:100 . When designing experiments, researchers should optimize antibody concentrations, with recommended dilution ranges of 1:500-2000 for WB and 1:20-200 for IHC-P applications .

How should researchers validate the specificity of IFRD2 antibodies?

Validation of IFRD2 antibody specificity is crucial for reliable experimental results. Researchers should implement multiple validation strategies including:

• Positive controls: Using cell lines known to express IFRD2, such as HeLa or PC-3 cells, as demonstrated in Western blot analyses .

• Negative controls: Employing IFRD2 knockout cells or tissues when available, similar to the IFRD2 knockout mice described in developmental studies .

• Band size verification: Confirming that the detected protein band is at the predicted molecular weight of 55 kDa for human IFRD2 .

• Cross-reactivity testing: Assessing potential cross-reactivity with the homologous IFRD1 protein, which shares significant sequence similarity, particularly in conserved domains .

• Peptide competition assays: Preincubating the antibody with excess immunizing peptide to confirm signal specificity.

These validation steps ensure that experimental results accurately represent IFRD2 expression and localization rather than non-specific binding or cross-reactivity.

What tissue expression patterns should researchers expect when using IFRD2 antibodies?

IFRD2 is expressed in various human tissues with notably higher expression in brain and muscle tissues . In experimental analyses, researchers should expect differential expression patterns across tissues, with significant expression also detected in the liver, kidney, heart, lung, and placenta . In fish models such as red sea bream (Pagrus major), IFRD2 shows particularly high expression in head kidney compared to trunk kidney (82.42-fold higher) . When conducting immunohistochemical analyses on human samples, positive staining has been documented in placental tissue . Researchers should anticipate that expression patterns may vary between species, developmental stages, and under different physiological or pathological conditions. For comparative studies across different species, it's important to consider sequence homology, which ranges from approximately 92.64% (large yellow croaker to red sea bream) to 53.85% (mouse to red sea bream) .

How can researchers optimize IFRD2 antibody detection in co-immunoprecipitation studies investigating ribosomal interactions?

Optimizing co-immunoprecipitation (Co-IP) protocols for studying IFRD2's ribosomal interactions requires special consideration of its mechanistic function. Since IFRD2 acts as a ribosome-binding protein that associates with the P- and E-sites of the ribosome and inserts a C-terminal helix into the mRNA exit channel , researchers should:

• Use gentle cell lysis conditions to preserve ribosomal complexes (e.g., low detergent concentrations).

• Include RNase inhibitors in buffers to maintain RNA-dependent interactions.

• Consider crosslinking approaches to stabilize transient interactions, as IFRD2-ribosome binding may be dynamic.

• Use sequential immunoprecipitation with antibodies against both IFRD2 and ribosomal markers (e.g., ribosomal proteins) to confirm co-complexes.

• Perform density gradient fractionation prior to immunoprecipitation to isolate polysome, monosome, and free ribosomal subunit fractions.

• Include appropriate controls using IFRD2 knockout cells (as generated in mouse studies ) to validate specificity.

For quantitative analysis of IFRD2-ribosome interactions, researchers can combine Co-IP with methods such as mass spectrometry or use proximity ligation assays to confirm in situ interactions.

What technical challenges might researchers encounter when using IFRD2 antibodies for immunohistochemistry in different tissue types?

Researchers applying IFRD2 antibodies in immunohistochemistry across diverse tissue types may encounter several technical challenges:

• Fixation sensitivity: IFRD2 epitopes may be differentially affected by fixation methods. While paraffin-embedded tissue protocols have been validated , researchers should optimize fixation time and conditions for each tissue type.

• Background signal: Polyclonal IFRD2 antibodies may produce higher background in tissues with abundant translation machinery. Implement stringent blocking steps (e.g., with 5-10% serum matching the secondary antibody species) and optimize antibody dilutions (starting with the recommended 1:20-200 range ).

• Tissue-specific expression levels: Given IFRD2's variable expression across tissues, with higher levels in brain and muscle , staining protocols may require adjustment for tissues with lower expression.

• Cross-reactivity with IFRD1: Due to the high sequence homology between IFRD1 and IFRD2 (reported 58% identity at cDNA level and 88% at amino acid level in mice ), antibody cross-reactivity is a significant concern. Validate antibody specificity using IFRD1/2 knockout tissues or with peptide competition.

• Distinguishing subcellular localization: As a ribosome-binding protein, IFRD2 may show cytoplasmic distribution patterns that can be difficult to distinguish from background. Consider counterstaining with ribosomal markers and using confocal microscopy for co-localization studies.

For tissues with complicated matrix components or high autofluorescence (like brain), tyramide signal amplification methods may improve specific signal detection while maintaining acceptable background levels.

How should researchers design experiments to investigate IFRD2's role in adipogenesis using antibody-based approaches?

Designing experiments to investigate IFRD2's role in adipogenesis requires multiple complementary approaches based on the established relationship between IFRD2 and adipocyte differentiation:

• Temporal expression profiling: Researchers should monitor IFRD2 protein levels during adipocyte differentiation using Western blot analysis at multiple time points. Evidence suggests IFRD2 functions as a translational inhibitor affecting adipogenesis through regulation of Dlk1 protein levels .

• Co-immunoprecipitation studies: Design Co-IP experiments to identify IFRD2 interaction partners in pre-adipocytes and during differentiation, particularly focusing on proteins involved in translational regulation and Wnt signaling components, as Wnt signaling is significantly altered in IFRD2-deficient models .

• Ribosome profiling with IFRD2 immunoprecipitation: Combine ribosome profiling with IFRD2 immunoprecipitation to identify mRNAs specifically regulated by IFRD2, focusing on adipogenic factors. This approach is supported by evidence that IFRD2 knockout increases polyribosome-bound Dlk1 mRNA levels .

• Immunocytochemistry during differentiation: Track IFRD2 subcellular localization changes during adipocyte differentiation, particularly in relation to markers of the translational machinery, as IFRD2 has been identified as a novel ribosome-binding protein .

• Protein synthesis assays: Compare global protein synthesis rates between wild-type and IFRD2-knockout or knockdown pre-adipocytes using 35S-methionine incorporation, as IFRD2 deficiency significantly increases general translational activity in fibroblasts .

When interpreting results, researchers should consider the potential functional redundancy between IFRD1 and IFRD2, as both contribute to adipogenic regulation but through different mechanisms: IFRD1 acts by controlling Wnt signaling, while IFRD2 functions as a translational inhibitor .

What control systems should be implemented when studying IFRD2 expression changes during immune challenges?

When investigating IFRD2 expression changes during immune challenges, researchers should implement comprehensive control systems based on observed patterns in fish models, where IFRD2 shows differential expression following pathogen exposure :

• Temporal controls:

  • Multiple time points should be analyzed (e.g., 1 hour, 12 hours, 1 day, 3 days, and 7 days post-infection) as IFRD2 shows time-dependent expression changes after pathogen challenge .

  • Include early time points (≤1 hour) to capture immediate early responses and extended time points (≥7 days) for resolution phase analysis.

• Tissue-specific controls:

  • Multiple tissues should be examined simultaneously, as IFRD2 expression responses are tissue-specific, with different patterns observed in gill, liver, kidney, and spleen tissues .

  • Include both immune-relevant tissues (spleen, kidney) and barrier tissues (gill, skin) to capture systemic and local responses.

• Pathogen-specific controls:

  • Different pathogens elicit distinct IFRD2 expression patterns. Compare responses to bacterial (e.g., E. piscicida, S. iniae) and viral (e.g., RSIV) challenges .

  • Include both Gram-positive and Gram-negative bacterial controls when studying bacterial responses.

• Dose-response controls:

  • Test multiple pathogen concentrations to determine if IFRD2 expression changes are dose-dependent.

  • Include sub-lethal and lethal challenge doses to differentiate between protective and pathological responses.

• Mock-infection controls:

  • Include vehicle-only controls matching all experimental handling procedures to account for stress-induced expression changes.

• Genetic controls:

  • When possible, include IFRD1 expression analysis as a reference for comparing related family members with potentially overlapping functions .

Implementation of these control systems will enable researchers to distinguish between specific immune response patterns and non-specific stress responses, providing more reliable interpretation of IFRD2's role in immunity.

How can researchers utilize IFRD2 antibodies in studies investigating translation regulation during cellular stress?

IFRD2 antibodies can be strategically employed to investigate translation regulation during cellular stress, capitalizing on IFRD2's established role as a ribosome-binding protein that inhibits mRNA translation :

• Stress granule co-localization: Researchers should employ dual immunofluorescence with IFRD2 antibodies and stress granule markers (e.g., G3BP1, TIA-1) across various stress conditions (oxidative stress, heat shock, ER stress). This approach reveals whether IFRD2 is recruited to stress granules where translation is temporarily halted.

• Polysome profiling with IFRD2 immunoblotting: Perform sucrose gradient fractionation of cytoplasmic lysates from stressed and unstressed cells, followed by Western blot analysis of fractions with IFRD2 antibodies. This technique can document stress-induced redistribution of IFRD2 between actively translating polysomes and non-translating mRNPs.

• Proximity-dependent biotin labeling: Use BioID or APEX2 fusions with IFRD2 to identify stress-specific interaction partners, providing insight into dynamic interactome changes during stress conditions.

• Phosphorylation-specific IFRD2 antibodies: Since many translation regulators are modified post-translationally during stress, developing phospho-specific IFRD2 antibodies can help determine if stress-activated kinases regulate IFRD2 function.

• IFRD2 immunoprecipitation coupled with RNA sequencing: This approach can identify specific mRNAs associated with IFRD2 during stress conditions, potentially revealing transcript-specific regulation patterns.

• Translational efficiency measurement: Combine polysome profiling with IFRD2 knockdown/overexpression to determine how IFRD2 levels influence the translation of specific mRNAs during stress, building on observations that IFRD2 affects general translational activity .

These approaches collectively can provide mechanistic insight into how IFRD2 contributes to stress-induced translational reprogramming, which is crucial for cellular adaptation and survival during adverse conditions.

What experimental approaches would resolve contradictions in IFRD2 tissue expression patterns across different species?

To resolve contradictions in IFRD2 tissue expression patterns across species, researchers should implement a multi-faceted experimental approach:

• Standardized cross-species expression analysis:

  • Perform simultaneous quantitative RT-PCR across equivalent tissues from multiple species using species-specific primers targeting conserved regions.

  • Normalize expression data to multiple reference genes validated for cross-species comparison.

  • This approach can quantitatively compare relative expression levels, similar to the 82.42-fold expression difference observed between head kidney and trunk kidney in red sea bream .

• Antibody validation for cross-species reactivity:

  • Test commercial IFRD2 antibodies against recombinant IFRD2 proteins from multiple species.

  • Generate custom antibodies against highly conserved epitopes for cross-species studies.

  • For each species, validate antibody specificity using tissue from IFRD2 knockout models when available .

• Immunohistochemical mapping:

  • Perform systematic IHC analysis across tissue panels from different species using validated antibodies .

  • Implement dual immunofluorescence with cell-type-specific markers to resolve cell-specific expression within complex tissues.

  • Document subcellular localization patterns to identify potential functional differences.

• Developmental trajectory analysis:

  • Map expression across developmental stages in multiple species, as IFRD2 is associated with early hematopoiesis and hepatic development in mice .

  • Use lineage tracing combined with IFRD2 immunostaining to identify dynamic expression changes during organogenesis.

• Functional conservation testing:

  • Perform cross-species complementation experiments by expressing IFRD2 orthologs in IFRD2-knockout systems.

  • Measure translation inhibition activity using in vitro translation assays with IFRD2 from different species.

• Pathogen challenge response comparison:

  • Challenge equivalent tissues from different species with identical pathogenic stimuli and compare IFRD2 expression responses .

  • This can reveal conserved versus species-specific immune regulatory functions.

By integrating these approaches, researchers can distinguish between true biological differences in IFRD2 tissue expression across species versus technical or methodological artifacts.

How should researchers interpret unexpected molecular weight variants of IFRD2 in Western blot experiments?

When encountering unexpected molecular weight variants of IFRD2 in Western blot experiments, researchers should systematically analyze potential biological and technical explanations:

• Predicted vs. Observed Molecular Weight:

  • The predicted molecular weight of human IFRD2 is approximately 55 kDa . Deviations from this expected size require careful interpretation.

  • For significant deviations, confirm primary antibody specificity through peptide competition assays or using IFRD2 knockout samples as negative controls .

• Interpreting Higher Molecular Weight Bands:

  • Post-translational modifications: IFRD2 may undergo ubiquitination, SUMOylation, or phosphorylation, increasing its apparent molecular weight.

  • Dimerization/complexes: Incomplete denaturation may result in IFRD2-containing complexes. Test by increasing SDS concentration or boiling time in sample preparation.

  • Isoforms: Verify against known splice variants by comparing band patterns with transcript data or using isoform-specific antibodies.

• Interpreting Lower Molecular Weight Bands:

  • Proteolytic degradation: Include protease inhibitors in all extraction buffers and minimize sample processing time and temperature.

  • Alternative translation start sites: Review the transcript sequence for potential downstream in-frame start codons.

  • C-terminal processing: IFRD2 contains a C-terminal helix that inserts into the ribosomal mRNA exit channel ; this domain might undergo regulated processing.

• Methodology for Resolving Ambiguities:

  • Perform 2D electrophoresis (isoelectric focusing followed by SDS-PAGE) to separate charge variants.

  • Use mass spectrometry to identify the exact composition of unexpected bands.

  • Compare results across different cell types and tissue sources, as IFRD2 is expressed differently across tissues .

  • Test multiple IFRD2 antibodies targeting different epitopes to confirm band identity.

• Quantification Considerations:

  • When quantifying IFRD2 by Western blot, clearly document which bands are included in the analysis.

  • In comparative studies, ensure consistent inclusion/exclusion criteria for band analysis across all samples.

This systematic approach helps distinguish biologically relevant IFRD2 variants from technical artifacts, leading to more accurate data interpretation.

What methodological adjustments can improve IFRD2 detection sensitivity in tissues with low expression levels?

To enhance IFRD2 detection sensitivity in tissues with low expression levels, researchers should implement several methodological refinements:

• Sample preparation optimization:

  • Enrich for IFRD2-containing fractions using subcellular fractionation, focusing on cytoplasmic and ribosome-associated fractions, given IFRD2's role as a ribosome-binding protein .

  • Use protein concentration methods (e.g., TCA precipitation, methanol/chloroform precipitation) before loading samples for Western blot.

  • For tissue sections in IHC, optimize antigen retrieval conditions through systematic comparison of heat-induced epitope retrieval in different pH buffers (citrate pH 6.0 vs. EDTA pH 9.0).

• Signal amplification strategies:

  • For Western blot: Implement more sensitive detection systems such as chemiluminescent substrates with enhanced sensitivity or near-infrared fluorescent secondary antibodies.

  • For IHC: Apply tyramide signal amplification (TSA) or polymer-based detection systems which can increase sensitivity by 10-100 fold compared to conventional methods .

  • For immunofluorescence: Use quantum dots as fluorescent labels, which provide higher signal intensity and photostability.

• Antibody optimization:

  • Test concentration series beyond the standard recommendations (1:500-2000 for WB; 1:20-200 for IHC-P) to identify optimal conditions for low-expression tissues.

  • Extend primary antibody incubation times (overnight at 4°C or longer) to enhance binding to sparse antigens.

  • Consider using cocktails of multiple IFRD2 antibodies targeting different epitopes simultaneously.

• Background reduction techniques:

  • Implement stringent blocking protocols using both protein blockers (e.g., BSA, casein) and serum matching the secondary antibody species.

  • Include detergents (0.1-0.3% Triton X-100 or Tween-20) in antibody diluents to reduce non-specific binding.

  • For tissues with high endogenous biotin, use avidin/biotin blocking steps before applying biotinylated detection systems.

• Quantitative approach adjustments:

  • Employ digital image analysis with background subtraction algorithms for more accurate quantification of weak signals.

  • Consider more sensitive detection methods such as proximity ligation assay (PLA) which can detect single protein molecules.

  • Use quantitative immunoprecipitation followed by targeted mass spectrometry for absolute quantification in samples with very low IFRD2 expression.

Implementation of these methodological adjustments has successfully enhanced detection of low-abundance proteins in challenging tissue types and can significantly improve IFRD2 visualization in tissues where expression is minimal.

How can IFRD2 antibodies be used to investigate the functional relationship between IFRD1 and IFRD2 in translational regulation?

Investigating the functional relationship between IFRD1 and IFRD2 in translational regulation requires sophisticated antibody-based approaches that can distinguish between these homologous proteins while revealing their potential overlapping or distinct functions:

• Differential localization studies:

  • Perform dual immunofluorescence using validated antibodies against IFRD1 and IFRD2 to map their subcellular distribution patterns in various cell types.

  • Analysis should focus on co-localization with ribosomal markers, as IFRD2 has been identified as a ribosome-binding protein that inhibits translation .

  • Quantify the degree of co-localization using methods such as Pearson's correlation coefficient or Manders' overlap coefficient.

• Sequential chromatin immunoprecipitation (ChIP-reChIP):

  • Evidence suggests IFRD1 acts by controlling Wnt signaling and thereby transcriptional regulation, while IFRD2 functions as a translational inhibitor .

  • Use ChIP-reChIP with IFRD1 and IFRD2 antibodies to determine if they can simultaneously associate with the same chromatin regions involved in translational regulation.

• Comparative ribosome interaction analysis:

  • Perform ribosome immunoprecipitation followed by mass spectrometry to identify the specific ribosomal proteins that interact with IFRD1 versus IFRD2.

  • Map the binding sites using cross-linking immunoprecipitation (CLIP) methods to determine if they compete for the same ribosomal binding sites or have distinct binding preferences.

• Reciprocal co-immunoprecipitation studies:

  • Immunoprecipitate IFRD1 and probe for IFRD2 (and vice versa) to test for direct protein-protein interactions or shared complex membership.

  • Include RNase treatment controls to determine if any detected interactions are RNA-dependent.

• Translational activity measurement in single vs. double knockout systems:

  • Compare global protein synthesis rates using metabolic labeling (35S-methionine incorporation) in wild-type, IFRD1-knockout, IFRD2-knockout, and double-knockout cells .

  • Analyze polysome profiles in these genetic backgrounds to assess changes in translation efficiency.

• Temporal dynamics during stress responses:

  • Monitor IFRD1 and IFRD2 protein levels and localization during various cellular stresses known to affect translation (e.g., ER stress, nutrient deprivation).

  • Determine if they show coordinated or independent regulation during stress recovery phases.

These approaches can reveal whether IFRD1 and IFRD2 function independently, synergistically, or antagonistically in translational regulation, building on evidence that they utilize different mechanisms to influence adipocyte differentiation .

What are the key methodological considerations for using IFRD2 antibodies in examining its role in pathogen-induced immune responses?

When investigating IFRD2's role in pathogen-induced immune responses using antibody-based methods, researchers should address several critical methodological considerations:

• Species-specific optimization:

  • Different species show distinct IFRD2 expression patterns and responses to pathogens. Red sea bream (Pagrus major) IFRD2 shows significant upregulation in the gills at 12 hours post-infection with S. iniae and at 3 days post-infection with RSIV .

  • Validate antibody specificity for the specific research species, as homology between species varies significantly (e.g., 78.49% homology between zebrafish and red sea bream IFRD2) .

  • For cross-species studies, target antibodies to the most conserved epitopes.

• Tissue-specific protocols:

  • IFRD2 expression varies dramatically between tissues, with up to 82.42-fold higher expression in head kidney compared to trunk kidney in red sea bream .

  • Optimize fixation and permeabilization protocols for each tissue type, particularly for immune-relevant tissues such as spleen, kidney, and gills in fish or lymph nodes and bone marrow in mammals.

  • Include tissue-specific positive controls to establish baseline expression levels.

• Temporal sampling design:

  • IFRD2 expression changes following pathogen exposure show distinct temporal patterns depending on the pathogen and tissue. For example, liver expression increases at 1 hour and 12 hours post-infection with E. piscicida .

  • Implement a comprehensive time-course sampling strategy spanning immediate (minutes to hours) and extended (days to weeks) timeframes.

  • Consider using in vivo imaging with fluorescently labeled antibodies for real-time monitoring in accessible tissues.

• Pathogen-specific considerations:

  • Different pathogens elicit distinct IFRD2 expression patterns. S. iniae infection causes significant upregulation in gills, kidney, and liver at different timepoints, while RSIV primarily affects gill expression .

  • Include multiple pathogen types (bacterial, viral, fungal) to distinguish general inflammatory responses from pathogen-specific regulation.

  • Consider using inactivated pathogens alongside live pathogens to differentiate between active infection and pattern recognition responses.

• Signaling pathway integration:

  • Combine IFRD2 antibody detection with phospho-specific antibodies against key immune signaling proteins (e.g., NF-κB, STATs, IRFs) to place IFRD2 regulation within established immune pathways.

  • Implement multiplexed immunoassays to simultaneously measure multiple immune factors alongside IFRD2.

• Technical controls for immune contexts:

  • Include isotype controls matched to IFRD2 antibody concentrations to control for non-specific binding, which can be particularly problematic in inflamed tissues.

  • Consider the impact of Fc receptor expression on immune cells when interpreting immunostaining results.

  • Account for potential changes in reference gene expression during immune responses when normalizing quantitative data.

These methodological considerations enable more reliable interpretation of IFRD2's dynamic regulation during immune responses and its potential contribution to host defense mechanisms.

What comprehensive validation pipeline should researchers implement before using a new IFRD2 antibody in high-stakes experiments?

Before deploying a new IFRD2 antibody in critical experiments, researchers should implement this systematic validation pipeline:

• Initial specificity assessment:

  • Western blot analysis using positive control samples (HeLa or PC-3 cell lysates) to confirm detection of the expected ~55 kDa band.

  • Peptide competition assay with the immunizing peptide to verify signal specificity.

  • Testing on IFRD2 knockout samples where available (e.g., MEFs from IFRD2 KO mice) to confirm absence of signal.

• Cross-reactivity evaluation:

  • Test against recombinant IFRD1 protein to assess potential cross-reactivity, given the high sequence homology between IFRD1 and IFRD2 (88% amino acid identity in mice) .

  • Analyze samples from IFRD1 knockout models to confirm IFRD2-specific signals.

  • Sequence alignment analysis to predict potential cross-reactive epitopes.

• Application-specific validation:

  • For Western blot: Optimize sample preparation, loading amount, and antibody dilution (1:500-2000 recommended range) .

  • For IHC: Test multiple antigen retrieval methods and antibody dilutions (1:20-200 recommended range) across diverse tissue types.

  • For immunoprecipitation: Verify pull-down efficiency using Western blot and mass spectrometry identification.

• Sensitivity and dynamic range determination:

  • Perform dilution series of recombinant IFRD2 protein to establish detection limits.

  • Compare antibody performance across tissues with known differential expression (e.g., brain/muscle vs. other tissues) .

  • Test detection capability under conditions that alter IFRD2 expression, such as pathogen challenge .

• Reproducibility assessment:

  • Test multiple antibody lots for consistent performance.

  • Validate across multiple technical platforms (e.g., different imaging systems, detection methods).

  • Implement standardized protocols with quantitative quality control metrics.

• Functional validation:

  • Confirm that antibody-detected IFRD2 localization is consistent with its known function as a ribosome-binding protein .

  • Verify changes in antibody signal following experimental manipulations that should alter IFRD2 levels (e.g., IFRD2 overexpression, siRNA knockdown).

  • For functional studies, confirm that antibody binding does not interfere with IFRD2's translational inhibition function .

This comprehensive validation pipeline ensures that experimental results obtained with the new IFRD2 antibody are reliable, reproducible, and accurately reflect the biological reality of IFRD2 expression and function.

What are the most promising research applications for IFRD2 antibodies based on current understanding of IFRD2 biology?

Based on current understanding of IFRD2 biology, several promising research applications for IFRD2 antibodies emerge:

• Translational regulation studies:

  • IFRD2 antibodies can be instrumental in deciphering tissue-specific translational control mechanisms, as IFRD2 functions as a ribosome-binding protein that inhibits mRNA translation .

  • Applications include ribosome immunoprecipitation followed by sequencing (RIBO-seq) to identify transcripts specifically regulated by IFRD2 during normal development and in disease states.

  • Particularly valuable for studying translational reprogramming during cellular stress, given IFRD2's role in ribosome inactivation .

• Developmental biology applications:

  • IFRD2 antibodies can track expression dynamics during embryonic development, particularly in early hematopoiesis after gastrulation and in hepatic primordium development .

  • Multi-color immunofluorescence with developmental markers can map IFRD2 expression in progenitor populations during lineage specification.

  • Particularly valuable for understanding developmental disorders affecting tissues with high IFRD2 expression, such as brain and muscle .

• Adipogenesis and metabolic research:

  • IFRD2 antibodies are crucial tools for investigating adipocyte differentiation regulation, as IFRD2 knockout mice show severely reduced adipose tissue and resistance to high-fat diet-induced obesity .

  • Applications include tracking IFRD2 expression during adipocyte differentiation and in metabolic disorders.

  • Particularly valuable for studying the interplay between translational control and adipogenic transcription factor networks.

• Comparative immunology:

  • IFRD2 antibodies can illuminate evolutionary conservation of immune regulatory mechanisms across species, as IFRD2 shows differential expression following pathogen challenges in fish models .

  • Applications include mapping IFRD2 responses to various pathogens across diverse vertebrate species.

  • Particularly valuable for understanding conserved versus species-specific innate immune regulatory pathways.

• Cancer biology:

  • IFRD2 antibodies can evaluate its potential role in cancer development and progression, particularly in tissues where it is highly expressed.

  • Applications include tumor tissue microarray analysis to correlate IFRD2 expression with clinical outcomes.

  • Particularly valuable for investigating translational control mechanisms in cancer cells, which often exhibit altered protein synthesis regulation.

• Neurodevelopmental research:

  • Given IFRD2's high expression in brain tissue , antibodies can map its neuroanatomical distribution and potential role in neurodevelopment.

  • Applications include co-localization studies with neuronal/glial markers to identify IFRD2-expressing neural cell populations.

  • Particularly valuable for understanding translational control in neuronal maturation and synaptic plasticity.