C22orf39 Antibody

What is C22orf39 and what cellular functions has it been associated with?

C22orf39 (Chromosome 22 Open Reading Frame 39) is also known as UPF0545 protein C22orf39. While initially classified as a protein of unknown function, research has begun to characterize its cellular role. The protein is encoded by a gene located on chromosome 22 and has several homologs across species, suggesting evolutionary conservation of function .

At the protein level, C22orf39 is designated as "UPF0545 protein C22orf39" or "UPF0545 protein C22orf39 homolog," with the UPF prefix indicating an uncharacterized protein family . Multiple alternative names exist across species, including RIKEN cDNA 2510002D24 gene in mouse models and chromosome specific homologs such as C22H22orf39 and C17H22orf39 in other organisms .

What types of C22orf39 antibodies are available for research applications?

Several types of C22orf39 antibodies are available for research, with key differences in:

• Target epitope specificity: Antibodies targeting specific amino acid regions such as AA 1-142

• Host species: Primarily rabbit-derived polyclonal antibodies for research applications

• Reactivity profile: Predominantly human-reactive, with cross-reactivity potential

• Antibody format: Unconjugated formats suitable for various detection methods

The choice of antibody depends on experimental design requirements and the specific C22orf39 region being investigated. Polyclonal antibodies provide broader epitope recognition, though monoclonal options may offer higher specificity for particular applications.

What are the validated applications for C22orf39 antibodies in research?

Current validation data supports using C22orf39 antibodies in:

• Enzyme-Linked Immunosorbent Assay (ELISA): Quantitative detection in solution-based formats

• Immunohistochemistry (IHC): Detection in tissue sections, particularly with antibodies such as ABIN7175041

• Western Blot (WB): Potential application, though additional validation may be required

While these represent the core validated applications, researchers should consider performing additional validation when applying these antibodies to novel experimental conditions or tissue types.

How might C22orf39 antibodies be utilized in cancer research studies?

Recent research indicates potential relevance of C22orf39 in cancer biology, particularly through alternative splicing mechanisms. Studies have identified alternative splicing events in C22orf39 (specifically C22orf39|61054|AT and C22orf39|61055|AT) that correlate with metastatic potential in endometrial cancer .

For cancer researchers, C22orf39 antibodies can be employed to:

• Evaluate expression patterns across normal versus malignant tissues

• Investigate correlations between protein expression and alternative splicing events

• Assess potential associations with metastasis biomarkers

• Explore functional relationships with known cancer-related proteins

When designing such experiments, researchers should consider antibodies that can distinguish between potential splice variants or post-translational modifications of C22orf39.

What controls and validation steps are essential when working with C22orf39 antibodies?

Given the relative novelty of C22orf39 research, rigorous validation is critical:

• Positive controls: Use recombinant C22orf39 protein or cells with confirmed expression

• Negative controls: Include isotype controls and tissues/cells known to lack C22orf39 expression

• Peptide competition assays: Confirm specificity by pre-incubating antibody with immunizing peptide

• Knockout/knockdown validation: Where possible, validate using CRISPR knockout or siRNA knockdown models

• Cross-reactivity assessment: Test against related proteins, particularly when investigating homologs

Antibody validation should be considered an ongoing process throughout a research project rather than a one-time event.

How do alternative splicing events affect C22orf39 antibody selection and experimental design?

Alternative splicing events in C22orf39 have been reported in cancer studies, with variants C22orf39|61054|AT and C22orf39|61055|AT showing significant correlations with metastatic potential . This has important implications for antibody selection:

• Epitope location assessment: Determine whether your antibody's epitope spans potential splice junctions

• Isoform specificity: Select antibodies that either specifically recognize or deliberately avoid splice variants

• Multiple antibody approach: Consider using antibodies targeting different regions to detect potential isoform diversity

When designing experiments investigating alternative splicing:

• Pair protein-level detection with transcript analysis (RT-PCR, RNA-seq)

• Consider using epitope tags in overexpression systems to track specific isoforms

• Document exact antibody clone/catalog information in publications to enhance reproducibility

What methodological optimizations improve C22orf39 detection in immunohistochemistry applications?

For optimal immunohistochemical detection of C22orf39:

• Antigen retrieval optimization: Test both heat-induced (citrate, EDTA) and enzymatic methods

• Blocking optimization: Extended blocking (3-5% BSA or serum) may reduce background

• Primary antibody concentration: Titration experiments should establish optimal dilution

• Incubation conditions: Extended overnight incubation at 4°C often improves signal-to-noise ratio

• Detection system selection: Polymer-based systems may offer enhanced sensitivity for low-abundance targets

The specific antibody ABIN7175041 has been validated for IHC applications, making it a potential candidate for such studies .

What are common technical challenges when working with C22orf39 antibodies and how can they be addressed?

Researchers often encounter these challenges when working with C22orf39 antibodies:

• Low signal intensity:

  • Increase antibody concentration

  • Extend incubation time

  • Use signal amplification systems

  • Optimize sample preparation protocols

• High background:

  • Increase blocking time/concentration

  • Test alternative blocking agents

  • Reduce primary antibody concentration

  • Include detergents in wash buffers

• Inconsistent results:

  • Standardize sample preparation

  • Establish positive control samples

  • Prepare larger antibody aliquots to reduce freeze-thaw cycles

  • Document lot-to-lot variation

How should researchers select between different C22orf39 antibodies for specific experimental applications?

Selection criteria should include:

• Target epitope: For studying specific domains or splice variants, select antibodies targeting relevant regions

• Validation data: Prioritize antibodies with validation in your specific application

• Host species compatibility: Consider downstream application requirements and secondary antibody availability

• Clonality: Polyclonal antibodies offer broader epitope recognition, while monoclonals provide consistency

• Species reactivity: Ensure compatibility with your experimental model system

For human research applications, antibodies like ABIN7175041, ABIN7175043, and ABIN7175044 offer documented reactivity, while studies in other species might utilize antibodies reactive with bovine, primate, or amphibian C22orf39 .

Is there evidence linking C22orf39 to specific disease pathways or potential therapeutic targets?

Current evidence suggests potential connections between C22orf39 and disease pathways:

• Cancer biology: Alternative splicing events in C22orf39 correlate with metastatic potential in endometrial cancer

• Potential co-expression relationships: Analysis has identified correlations between C22orf39 alternative transcripts and genes involved in cancer progression

These observations warrant further investigation into:

• Mechanistic studies of C22orf39 function in cellular pathways

• Comprehensive expression profiling across normal and disease tissues

• Potential protein-protein interactions with known disease mediators

How can C22orf39 antibodies be integrated into multi-omics research approaches?

Modern multi-omics approaches can leverage C22orf39 antibodies in several ways:

• Proteogenomic integration:

  • Correlate protein expression (via antibody-based methods) with transcriptomic analysis of splice variants

  • Combine with genomic data to identify regulatory mechanisms and genetic associations

• Spatial biology:

  • Apply C22orf39 antibodies in multiplexed immunofluorescence or imaging mass cytometry

  • Correlate with spatial transcriptomics data to map expression patterns at tissue level

• Interactome analysis:

  • Use antibodies for co-immunoprecipitation followed by mass spectrometry

  • Identify binding partners and protein complexes containing C22orf39

• Functional genomics:

  • Validate genetic screen hits using antibody-based validation

  • Correlate protein expression changes with cellular phenotypes

What is the recommended protocol for using C22orf39 antibodies in immunohistochemistry?

Based on validated antibody applications, the following protocol is recommended:

• Sample preparation:

  • Fix tissues in 10% neutral buffered formalin (24 hours)

  • Process and embed in paraffin

  • Section at 4-6 μm thickness

• Deparaffinization and rehydration:

  • Xylene: 2 × 5 minutes

  • 100% ethanol: 2 × 3 minutes

  • 95% ethanol: 1 × 3 minutes

  • 70% ethanol: 1 × 3 minutes

  • Distilled water: 5 minutes

• Antigen retrieval:

  • Heat-induced epitope retrieval in citrate buffer (pH 6.0)

  • Maintain at 95-98°C for 20 minutes

  • Cool to room temperature (20 minutes)

• Blocking and antibody incubation:

  • Block endogenous peroxidase: 3% H₂O₂ (10 minutes)

  • Protein block: 5% normal goat serum (1 hour)

  • Primary antibody: Anti-C22orf39 (1:100 dilution in antibody diluent)

  • Incubate overnight at 4°C in humidified chamber

• Detection and visualization:

  • Wash: PBS with 0.1% Tween-20 (3 × 5 minutes)

  • Secondary antibody: HRP-conjugated anti-rabbit (30 minutes)

  • Wash: PBS with 0.1% Tween-20 (3 × 5 minutes)

  • Develop: DAB substrate (monitor for 2-10 minutes)

  • Counterstain: Hematoxylin (30 seconds)

  • Dehydrate and mount with permanent mounting medium

This protocol may require optimization based on specific tissue types and fixation conditions.

How can researchers effectively troubleshoot non-specific binding when using C22orf39 antibodies?

When encountering non-specific binding:

• Systematic optimization steps:

  • Titrate primary antibody concentration (1:50 to 1:500 dilution series)

  • Extend blocking time (2-16 hours)

  • Test alternative blocking agents (BSA, casein, commercial blockers)

  • Increase detergent concentration in wash buffers (0.1-0.3% Tween-20)

• Advanced troubleshooting approaches:

  • Pre-adsorb antibody with cell/tissue lysate lacking target

  • Perform peptide competition assays

  • Test alternative antibody clones targeting different epitopes

  • Implement post-fixation steps to reduce background

• Assay-specific adjustments:

  • For IHC: Optimize DAB development time and quenching steps

  • For IF: Include additional autofluorescence quenching steps

  • For ELISA: Consider sandwich format instead of direct coating