AKR1E2 Antibody

What is AKR1E2 and what are its primary biological functions?

AKR1E2 (Aldo-Keto Reductase Family 1, Member E2) is a cytoplasmic enzyme that functions primarily as a 1,5-anhydro-D-fructose reductase. The protein catalyzes the NADPH-dependent reduction of 1,5-anhydro-D-fructose (AF) to 1,5-anhydro-D-glucitol. Additionally, AKR1E2 demonstrates low NADPH-dependent reductase activity toward 9,10-phenanthrenequinone in vitro, suggesting potential roles in detoxification pathways . This enzyme belongs to the wider aldo/keto reductase superfamily, sharing structural and functional characteristics with other AKR proteins involved in various metabolic processes . Its highly tissue-specific expression pattern suggests specialized functions in testicular biology that warrant further investigation through antibody-based research techniques.

What is the expression profile of AKR1E2 in human tissues?

AKR1E2 demonstrates a highly tissue-specific expression pattern, being predominantly expressed in the testis according to multiple independent studies . More specifically, the protein is localized within testicular germ cells and testis interstitial cells . This restricted expression profile suggests AKR1E2 may play specialized roles in male reproductive physiology or spermatogenesis. When designing experiments to study AKR1E2, researchers should select appropriate positive controls (testicular tissue) and negative controls (non-testicular tissues) to validate antibody specificity. The cytoplasmic localization of AKR1E2 further informs appropriate cellular fractionation techniques when preparing samples for antibody-based detection methods.

What are the molecular characteristics of AKR1E2 that influence antibody selection?

AKR1E2 has a calculated molecular weight of 36,589 Da , which researchers should consider when validating antibody specificity by Western blot. The protein is also known by several alternative names including AKR1CL2, AKRDC1, HTSP1, LoopADR, TAKR, and hTSP . When selecting antibodies, researchers should verify which protein epitopes are targeted, as some commercially available antibodies are developed against the C-terminal region (amino acids 291-320) while others may target different regions. Understanding the protein's tertiary structure and post-translational modifications is essential for selecting antibodies that target accessible epitopes in native versus denatured conditions.

What criteria should guide the selection of AKR1E2 antibodies for specific applications?

When selecting an AKR1E2 antibody, researchers should consider:

For Western blot applications, polyclonal antibodies targeting the C-terminal region (amino acids 291-320) of AKR1E2 have been validated . These antibodies are purified through protein A columns followed by peptide affinity purification to ensure specificity . For immunohistochemistry, researchers should consider antibodies validated specifically for fixed tissue samples and optimize protocols for testicular tissue, where AKR1E2 is predominantly expressed .

How can I validate the specificity of an AKR1E2 antibody in my experimental system?

A comprehensive validation strategy for AKR1E2 antibodies should include:

• Positive and negative tissue controls: Use testicular tissue (positive) versus non-testicular tissues (negative) based on the known expression profile .

• Peptide competition assay: Pre-incubate the antibody with the immunizing peptide before application to verify signal extinction.

• Molecular weight verification: Confirm detection of a band at approximately 36.6 kDa in Western blot applications .

• Knockout/knockdown controls: If available, use AKR1E2 knockout models or siRNA-mediated knockdown samples as negative controls.

• Multiple antibody comparison: Use antibodies targeting different epitopes of AKR1E2 to corroborate findings.

For IHC applications, include peptide blocking controls and compare staining patterns with literature reports describing testicular germ cells and interstitial cell localization . For ELISA applications, establish a standard curve using recombinant AKR1E2 protein and verify detection sensitivity within the expected range (5.0-100 ng/mL) .

What are the optimal storage conditions for maintaining AKR1E2 antibody performance?

To maintain optimal antibody performance, store AKR1E2 antibodies according to manufacturer recommendations. Generally, antibodies should be stored at -20°C for long-term storage, with small aliquots to prevent freeze-thaw cycles that can degrade antibody quality . For short-term use (up to one month), storage at 4°C is acceptable . Most commercial AKR1E2 antibodies are supplied in PBS containing preservatives such as 0.09% sodium azide or 50% glycerol with 0.5% BSA . When working with these antibodies, researchers should document lot numbers and maintain consistent storage conditions throughout a research project to minimize variability. Additionally, avoid exposing antibodies to direct light and minimize exposure to room temperature, as these conditions can accelerate protein degradation.

What are the recommended protocols for Western blot detection of AKR1E2?

For optimal Western blot detection of AKR1E2:

• Sample preparation:

  • Extract proteins from testicular tissue or cells using RIPA buffer supplemented with protease inhibitors

  • Determine protein concentration using BCA or Bradford assay

  • Load 20-50 μg total protein per lane

• Electrophoresis conditions:

  • Use 10-12% SDS-PAGE gels

  • Include molecular weight markers spanning 25-50 kDa range

• Transfer and blocking:

  • Transfer to PVDF membrane at 100V for 60-90 minutes

  • Block with 5% non-fat milk in TBST for 1 hour at room temperature

• Antibody incubation:

  • Dilute primary AKR1E2 antibody 1:1000 in blocking buffer

  • Incubate overnight at 4°C with gentle rocking

  • Wash 3× with TBST, 10 minutes each

  • Incubate with HRP-conjugated secondary antibody (1:5000) for 1 hour at room temperature

• Detection:

  • Develop using ECL substrate

  • Expose to X-ray film or image using digital system

  • Verify band at approximately 36.6 kDa

For validation purposes, include positive control (testis lysate) and negative control (non-testicular tissue) samples . If multiple bands appear, consider using peptide competition assays to verify specificity.

How should I optimize immunohistochemistry protocols for AKR1E2 detection in testicular tissues?

For immunohistochemical detection of AKR1E2 in testicular tissues:

• Tissue preparation:

  • Fix tissues in 10% neutral-buffered formalin for 24 hours

  • Process and embed in paraffin

  • Section at 4-5 μm thickness

• Antigen retrieval:

  • Perform heat-induced epitope retrieval using citrate buffer (pH 6.0)

  • Heat in pressure cooker or microwave for 15-20 minutes

  • Cool sections to room temperature (approximately 20 minutes)

• Blocking and antibody incubation:

  • Block endogenous peroxidase with 3% H₂O₂ for 10 minutes

  • Block non-specific binding with 5% normal goat serum for 1 hour

  • Apply AKR1E2 antibody at 1:100-1:300 dilution overnight at 4°C

  • Wash 3× with PBS, 5 minutes each

• Detection and visualization:

  • Apply HRP-conjugated secondary antibody for 1 hour at room temperature

  • Develop with DAB substrate

  • Counterstain with hematoxylin

  • Dehydrate, clear, and mount

Always include positive control (normal testis) and negative controls (primary antibody omission and non-testicular tissue) . Optimize antibody concentration specifically for your tissue samples and fixation conditions. Focus analysis on cytoplasmic staining in testicular germ cells and interstitial cells, consistent with the known localization of AKR1E2 .

What considerations are important when designing ELISA-based experiments for AKR1E2 quantification?

When implementing ELISA for AKR1E2 quantification:

• ELISA format selection:

  • Competition ELISA is recommended for AKR1E2 detection

  • Detection range: 5.0-100 ng/mL

  • Minimum detection limit: 5.0 ng/mL

  • Sensitivity: 1.0 ng/mL

• Sample preparation:

  • Compatible sample types: cell culture supernatant, plasma, serum, tissue homogenate

  • Prepare tissue homogenates in PBS with protease inhibitors

  • Centrifuge at 3000 × g to remove particulates

  • Dilute samples appropriately to fall within standard curve range

• Standard curve preparation:

  • Use the provided standards (A-F) to generate a standard curve

  • Plot concentration versus optical density

  • Prepare fresh standards for each assay

• Antibody dilution:

  • Dilute AKR1E2 antibody 1:10,000 for ELISA applications

  • Optimize dilution based on signal-to-noise ratio

• Controls and validation:

  • Include blank wells (no sample)

  • Run samples in duplicate or triplicate

  • Include positive control (testis extract) and negative control (non-testicular tissue)

For quantitative analysis, ensure all equipment is properly calibrated, particularly the microplate reader measuring absorbance at 450 nm . Analytical software should incorporate a 4- or 5-parameter logistic curve fit for optimal quantification accuracy.

How can AKR1E2 antibodies be utilized to investigate protein-protein interactions?

For investigating AKR1E2 protein interactions:

• Co-immunoprecipitation (Co-IP):

  • Prepare testicular cell or tissue lysates in non-denaturing buffer

  • Pre-clear lysate with Protein A/G beads for 1 hour

  • Incubate cleared lysate with AKR1E2 antibody overnight at 4°C

  • Add Protein A/G beads and incubate for 2-4 hours

  • Wash beads 4-5 times with wash buffer

  • Elute bound proteins and analyze by Western blot using antibodies against suspected interacting partners

• Proximity Ligation Assay (PLA):

  • Fix cells or tissue sections and permeabilize

  • Block non-specific binding sites

  • Incubate with AKR1E2 antibody and antibody against putative interacting protein

  • Follow manufacturer's protocol for PLA probes and detection

  • Analyze fluorescent signals indicating proteins in close proximity (<40 nm)

• Pull-down assays:

  • Express and purify recombinant AKR1E2 with appropriate tag

  • Immobilize on matrix or beads

  • Incubate with cellular lysates

  • Wash, elute, and identify binding partners by mass spectrometry

Focus investigations on potential interactions with NADPH-dependent enzymes or proteins involved in testicular metabolism, considering AKR1E2's enzymatic function and tissue-specific expression . Use appropriate negative controls including IgG controls and samples from tissues that do not express AKR1E2.

What approaches can be used to study AKR1E2 enzymatic activity in conjunction with antibody-based detection?

To study AKR1E2 enzymatic activity alongside antibody detection:

• Activity assays following immunoprecipitation:

  • Immunoprecipitate AKR1E2 from testicular lysates using validated antibodies

  • Measure NADPH-dependent reduction of 1,5-anhydro-D-fructose to 1,5-anhydro-D-glucitol

  • Monitor NADPH consumption by spectrophotometric methods (decrease in absorbance at 340 nm)

  • Correlate activity with protein levels determined by Western blot

• In situ activity assays combined with immunohistochemistry:

  • Perform enzymatic activity assay on tissue sections

  • Follow with immunohistochemical detection of AKR1E2

  • Compare patterns of activity with protein localization

• Structure-function analysis:

  • Generate AKR1E2 mutants affecting catalytic sites

  • Express in appropriate cell systems

  • Correlate antibody detection of protein levels with enzymatic activity

  • Use to identify critical residues for catalysis

• Inhibitor studies:

  • Test effects of potential inhibitors on AKR1E2 activity

  • Verify target engagement using antibody-based methods

  • Correlate inhibition with structural interactions

When designing these experiments, researchers should consider the optimal buffer conditions for AKR1E2 activity, including pH, temperature, and cofactor concentrations. Control experiments should include enzyme kinetics characterization with recombinant AKR1E2 protein to establish baseline parameters.

How can AKR1E2 antibodies be applied in developmental studies of testicular biology?

For developmental studies of AKR1E2 in testicular biology:

• Temporal expression analysis:

  • Collect testicular tissues at different developmental stages

  • Perform Western blot analysis using AKR1E2 antibodies at 1:1000 dilution

  • Quantify expression relative to loading controls

  • Correlate with developmental markers

• Spatial expression patterns:

  • Conduct immunohistochemistry on testicular sections from different developmental stages

  • Use AKR1E2 antibodies at 1:100-1:300 dilution

  • Co-stain with markers for specific cell types (germ cells, Sertoli cells, Leydig cells)

  • Analyze changes in cellular localization during development

• Cell-type specific expression:

  • Isolate specific testicular cell populations using flow cytometry or magnetic separation

  • Verify AKR1E2 expression by Western blot and RT-PCR

  • Correlate protein levels with cell differentiation state

• Functional studies during development:

  • Design loss-of-function or gain-of-function experiments

  • Monitor effects on spermatogenesis and testicular development

  • Use AKR1E2 antibodies to confirm experimental manipulation effectiveness

These approaches should incorporate appropriate controls, including tissues where AKR1E2 is not expressed . Experimental design should account for the potential influence of hormonal regulation on AKR1E2 expression during developmental transitions, particularly during puberty and sexual maturation.

Why might Western blot analysis with AKR1E2 antibodies show unexpected banding patterns?

When troubleshooting unexpected Western blot banding patterns:

For AKR1E2 specifically, validate all bands by peptide competition assay using the synthetic peptide corresponding to amino acids 291-320 . Consider that AKR1E2 may interact with other proteins in the aldo-keto reductase family, potentially leading to cross-reactivity. Always compare your results with positive controls (testis lysate) where the expected 36.6 kDa band should be visible .

How should I interpret inconsistent results between detection methods for AKR1E2?

When facing inconsistencies between different detection methods:

• Western blot vs. IHC discrepancies:

  • Western blot detects denatured proteins while IHC detects native conformations

  • Epitope accessibility may differ between methods

  • Verify antibody performance in each application separately

  • Consider using multiple antibodies targeting different epitopes

• ELISA vs. Western blot inconsistencies:

  • ELISA sensitivity (1.0 ng/mL) may exceed Western blot detection limits

  • Competition ELISA format may be affected by interfering substances

  • Verify sample matrix effects with spike-recovery experiments

  • Consider sample concentration methods for Western blot

• General approach to resolving inconsistencies:

  • Systematically evaluate each method's controls

  • Verify antibody batch consistency and storage conditions

  • Consider third method validation (e.g., mass spectrometry)

  • Evaluate sample preparation effects on protein stability and detection

Document all methodological details, including antibody dilutions (1:500-1:2000 for WB; 1:100-1:300 for IHC; 1:10,000 for ELISA) and detection conditions to facilitate troubleshooting. Remember that different detection methods may reveal different aspects of AKR1E2 biology, potentially reflecting biologically relevant differences rather than methodological errors.

What are the critical controls needed when studying tissue-specific expression of AKR1E2?

For rigorous analysis of AKR1E2 tissue-specific expression:

• Essential tissue controls:

  • Positive control: Testicular tissue (known to express AKR1E2)

  • Negative controls: Multiple non-testicular tissues

  • Gradient controls: Tissues with potential low-level expression

• Antibody validation controls:

  • Peptide competition: Pre-incubate antibody with immunizing peptide

  • Isotype control: Use matched IgG at same concentration

  • Antibody omission: Perform protocol without primary antibody

• Method-specific controls:

  • Western blot: Recombinant AKR1E2 protein as positive control

  • IHC: Serial dilutions of primary antibody to demonstrate specificity

  • ELISA: Standard curve with recombinant protein (5.0-100 ng/mL range)

• Biological validation:

  • Correlate protein detection with mRNA expression (RT-PCR)

  • Confirm cellular localization (cytoplasmic) using subcellular fractionation

  • Consider developmental stage effects on expression

When interpreting results, researchers should acknowledge that very low expression levels might be detected by sensitive methods like ELISA (detection limit 5.0 ng/mL) but not by less sensitive techniques. Additionally, post-translational modifications may affect epitope recognition differently across tissues, potentially leading to false negatives. Using antibodies targeting different regions of AKR1E2 can help address this limitation.

How can I design experiments to investigate the role of AKR1E2 in male reproductive physiology?

For investigating AKR1E2's role in reproductive physiology:

• Functional genomics approaches:

  • CRISPR-Cas9 knockout of AKR1E2 in testicular cell lines

  • shRNA knockdown in primary testicular cultures

  • Validate knockdown/knockout using AKR1E2 antibodies at 1:1000 dilution

  • Assess phenotypic changes in cell morphology, proliferation, and differentiation

• Animal model studies:

  • Generate AKR1E2 knockout mouse models

  • Characterize reproductive phenotypes including:

    • Fertility parameters

    • Testicular histology

    • Spermatogenesis progression

    • Sperm function

  • Use AKR1E2 antibodies (1:100-1:300) for IHC validation of knockout

• Metabolomic analysis:

  • Compare 1,5-anhydro-D-fructose and 1,5-anhydro-D-glucitol levels in:

    • Wild-type vs. AKR1E2-deficient models

    • Different stages of spermatogenesis

    • Various testicular cell populations

  • Correlate metabolite levels with AKR1E2 protein expression

• Clinical correlations:

  • Analyze AKR1E2 expression in testicular biopsies from patients with:

    • Male infertility

    • Testicular cancer

    • Developmental disorders

  • Correlate expression with clinical parameters and outcomes

These experimental approaches should incorporate appropriate controls and validation using antibodies with confirmed specificity for AKR1E2. Researchers should consider the cytoplasmic localization of AKR1E2 when interpreting results and designing functional studies.

What techniques can be used to study post-translational modifications of AKR1E2?

To investigate post-translational modifications (PTMs) of AKR1E2:

• Phosphorylation analysis:

  • Immunoprecipitate AKR1E2 using specific antibodies

  • Analyze by Western blot using phospho-specific antibodies

  • Alternatively, use mass spectrometry to identify phosphorylation sites

  • Validate functional significance with phosphatase treatments

• Glycosylation studies:

  • Treat samples with deglycosylation enzymes (PNGase F, O-glycosidase)

  • Compare molecular weight shifts by Western blot

  • Use lectins to probe for specific glycan structures

  • Correlate glycosylation status with enzymatic activity

• Ubiquitination and SUMOylation:

  • Immunoprecipitate AKR1E2 under denaturing conditions

  • Probe with anti-ubiquitin or anti-SUMO antibodies

  • Use proteasome inhibitors to enhance detection

  • Correlate modifications with protein stability and turnover

• PTM site mapping:

  • Generate recombinant AKR1E2 mutants at predicted modification sites

  • Express in appropriate cell systems

  • Compare PTM patterns and functional consequences

  • Validate with site-specific antibodies if available

These approaches should include appropriate controls and consider the tissue-specific expression of AKR1E2 in testis . Mass spectrometry analyses should target the 36.6 kDa region and adjacent molecular weights to capture modified forms. Researchers should correlate PTMs with AKR1E2's NADPH-dependent reductase activity to establish functional significance.

How can multiplexed detection methods enhance AKR1E2 research in complex testicular samples?

For multiplexed detection of AKR1E2 in testicular samples:

• Multiplex immunofluorescence:

  • Combine AKR1E2 antibodies with markers for:

    • Cell type-specific markers (VASA for germ cells, SOX9 for Sertoli cells)

    • Developmental stage markers (SYCP3 for meiotic cells)

    • Functional pathway components (NADPH-producing enzymes)

  • Use spectrally distinct fluorophores and multispectral imaging

  • Perform quantitative colocalization analysis

• Mass cytometry (CyTOF):

  • Label AKR1E2 antibodies with rare earth metals

  • Combine with multiple markers for cellular phenotyping

  • Analyze testicular cell suspensions at single-cell resolution

  • Perform high-dimensional data analysis to identify cell populations

• Single-cell analysis workflows:

  • Sort testicular cells based on surface markers

  • Analyze AKR1E2 expression in specific populations by Western blot

  • Correlate with single-cell transcriptomics data

  • Identify regulatory relationships

• Spatial transcriptomics with protein detection:

  • Perform in situ hybridization for AKR1E2 mRNA

  • Follow with immunodetection of AKR1E2 protein

  • Analyze spatial relationships with other markers

  • Correlate mRNA and protein expression patterns

These multiplexed approaches should include appropriate controls for antibody specificity, including peptide competition controls and validation in tissues known to lack AKR1E2 expression. Consider the detection range (5.0-100 ng/mL) and sensitivity (1.0 ng/mL) when designing experiments to ensure signals remain within quantifiable ranges.

How can AKR1E2 antibodies be applied in biomarker development for testicular disorders?

For applying AKR1E2 as a potential biomarker:

• Tissue microarray analysis:

  • Construct tissue microarrays from various testicular pathologies

  • Perform immunohistochemistry using AKR1E2 antibodies (1:100-1:300)

  • Quantify expression patterns using digital pathology

  • Correlate expression with clinical parameters and outcomes

• Liquid biopsy development:

  • Develop sensitive ELISA protocols (detection limit 5.0 ng/mL)

  • Evaluate AKR1E2 levels in seminal plasma or blood

  • Compare levels between healthy individuals and those with testicular disorders

  • Assess predictive value for specific conditions

• Multimarker panel development:

  • Combine AKR1E2 detection with established testicular markers

  • Perform multivariate analysis to improve diagnostic accuracy

  • Validate in independent cohorts

  • Determine sensitivity and specificity for specific conditions

• Longitudinal monitoring protocols:

  • Establish baseline AKR1E2 levels in healthy individuals

  • Monitor changes during disease progression or treatment

  • Correlate with clinical response

  • Develop standardized testing protocols

These approaches should incorporate appropriate quality control measures, including standard curves using recombinant AKR1E2 protein and spike-recovery experiments to validate detection in complex biological matrices. Given AKR1E2's testis-specific expression , researchers should evaluate its potential as a tissue-specific marker for testicular damage or dysfunction.

What experimental strategies can reveal AKR1E2's role in enzymatic pathways relevant to testicular metabolism?

To investigate AKR1E2's role in testicular metabolic pathways:

• Metabolic substrate profiling:

  • Purify AKR1E2 using immunoprecipitation with specific antibodies

  • Screen against panel of potential substrates beyond known targets

  • Monitor NADPH consumption spectrophotometrically

  • Identify novel substrates relevant to testicular metabolism

• Metabolomic analysis in AKR1E2-manipulated systems:

  • Generate AKR1E2 knockdown/knockout models

  • Perform untargeted metabolomics

  • Focus on pathways involving 1,5-anhydro-D-fructose and related compounds

  • Validate findings with targeted metabolite quantification

• Pathway intersection analysis:

  • Use AKR1E2 antibodies for co-immunoprecipitation

  • Identify interacting proteins by mass spectrometry

  • Map interactions to known metabolic pathways

  • Validate functional relationships with enzymatic assays

• Flux analysis:

  • Use isotope-labeled substrates in testicular cells or tissues

  • Track metabolite conversions in presence/absence of AKR1E2

  • Correlate with protein expression levels

  • Define rate-limiting steps in relevant pathways

These approaches should include appropriate controls and consider AKR1E2's NADPH-dependent reductase activity . Researchers should validate findings across multiple experimental systems and correlate with the known cytoplasmic localization of AKR1E2 in testicular cells.

How should differences in antibody performance be addressed when comparing AKR1E2 data across studies?

To address antibody performance variability in cross-study comparisons:

• Standardization strategies:

  • Develop standard reference materials (recombinant AKR1E2)

  • Establish calibration curves across detection methods

  • Use consistent positive controls (testicular tissue lysates)

  • Report antibody catalog numbers, lot numbers, and validation data

• Cross-validation protocols:

  • Test multiple antibodies on identical samples

  • Compare detection limits and dynamic ranges

  • Establish conversion factors between different antibodies

  • Document epitope differences between antibodies

• Meta-analysis approach:

  • Standardize data reporting formats

  • Apply normalization methods to account for antibody differences

  • Use relative rather than absolute quantification when comparing studies

  • Weight studies by quality of antibody validation

• Reporting guidelines:

  • Document complete antibody characteristics:

    • Target epitope (e.g., C-terminal region, aa 291-320)

    • Antibody type (polyclonal/monoclonal)

    • Validation methods employed

    • Optimal dilutions for specific applications (1:500-1:2000 for WB; 1:100-1:300 for IHC)

Researchers should establish internal reference standards when comparing historical data and consider generating bridging datasets when transitioning between antibody reagents. When possible, validation with orthogonal methods such as mass spectrometry can provide antibody-independent confirmation of findings.

What emerging technologies could advance AKR1E2 antibody-based research?

Emerging technologies for AKR1E2 research include:

• Nanobody and recombinant antibody development:

  • Generate single-domain antibodies against AKR1E2

  • Engineer antibodies with enhanced specificity for distinct epitopes

  • Develop antibody fragments maintaining specificity but with improved tissue penetration

  • Create genetically encoded intrabodies for live-cell imaging

• Advanced imaging technologies:

  • Super-resolution microscopy of AKR1E2 localization

  • Live-cell imaging using fluorescent AKR1E2 antibody fragments

  • Correlative light and electron microscopy for ultrastructural localization

  • Expansion microscopy for enhanced spatial resolution

• Microfluidic and single-cell applications:

  • Develop microfluidic antibody-based capture systems

  • Combine with single-cell transcriptomics

  • Create droplet-based enzymatic activity assays

  • Enable high-throughput screening of AKR1E2 modulators

• Synthetic biology approaches:

  • Design split-protein systems based on AKR1E2 interactions

  • Create biosensors for AKR1E2 substrates and products

  • Develop optogenetic tools to control AKR1E2 activity

  • Engineer cellular reporters for AKR1E2 expression and function

These technologies should build upon current knowledge of AKR1E2's structure, tissue distribution, and enzymatic function . Researchers should validate new approaches against established methods using well-characterized antibodies with documented specificity and performance characteristics.

How might integrated multi-omics approaches enhance our understanding of AKR1E2 function?

For integrated multi-omics investigation of AKR1E2:

• Proteogenomic integration:

  • Correlate AKR1E2 protein levels (antibody-based detection) with:

    • Genomic variants affecting expression or function

    • Transcriptomic data from various testicular cell types

    • Epigenetic modifications affecting gene regulation

  • Identify regulatory mechanisms controlling tissue-specific expression

• Structural biology with functional validation:

  • Determine AKR1E2 crystal structure

  • Map epitopes recognized by various antibodies

  • Correlate structural features with enzymatic function

  • Design structure-based inhibitors and activators

• Systems biology modeling:

  • Integrate AKR1E2 into testicular metabolic network models

  • Predict metabolic flux changes under various conditions

  • Validate predictions using antibody-based quantification

  • Identify emergent properties from network analysis

• Spatially resolved multi-omics:

  • Combine immunodetection with spatial transcriptomics

  • Map AKR1E2 protein distribution relative to substrate availability

  • Correlate with local metabolite concentrations

  • Develop computational models of spatial regulation