C17orf47 Antibody

What is C17orf47 and its relationship to SEPTIN4?

C17orf47 is a reported synonym of the SEPTIN4 gene. The gene encodes septin 4, a protein that functions primarily in the apoptotic pathway among other biological roles. This relationship between C17orf47 and SEPTIN4 is critical for researchers to understand when selecting and using antibodies against this target . The gene has been assigned the GeneID 284083 and has the UniProt Primary Accession Number Q8NEP4, with a secondary accession number of Q8N821 . Understanding this nomenclature relationship helps researchers avoid confusion when searching literature and databases for information about this protein.

What are the key structural and functional characteristics of C17orf47/SEPTIN4 protein?

The human version of C17orf47/SEPTIN4 has a canonical amino acid length of 478 residues and a protein mass of approximately 55.1 kilodaltons . Eight different isoforms of the protein have been identified, which is important to consider when designing experiments . Subcellularly, the protein is reported to be localized in cytoplasmic vesicles and the cytoplasm of cells . Functionally, SEPTIN4 is known to play important roles in the apoptotic pathway, making it a significant target for research in cell death mechanisms and related pathological conditions . The protein's various domains and post-translational modifications should be considered when selecting antibodies for specific research applications.

What are the common research applications for C17orf47 antibodies?

C17orf47 antibodies are primarily used in several key research applications:

• ELISA (Enzyme-Linked Immunosorbent Assay): For quantitative detection of the target protein in biological samples .

• Immunohistochemistry (IHC): For visualizing the protein's expression and localization in tissue samples .

• Western Blot (WB): Some antibodies are validated for detecting C17orf47/SEPTIN4 in protein extracts .

• Immunocytochemistry (ICC): For cellular localization studies .

• Immunofluorescence (IF): For high-resolution imaging of the protein's distribution .

The selection of the appropriate application depends on your specific research question, the sample type, and the level of sensitivity required for detection.

What types of C17orf47 antibodies are available and how should they be selected?

Researchers have multiple options when selecting C17orf47 antibodies, including:

Most available antibodies are polyclonal, raised in rabbits against recombinant human C17orf47 protein . The selection should be based on the specific experimental requirements, including the detection method, sample type, and required sensitivity . For specialized applications like flow cytometry or ChIP assays, verify that the antibody has been validated for these specific uses before purchase.

How should C17orf47 antibodies be validated for experimental use?

Proper validation of C17orf47 antibodies is essential to ensure reliable experimental results:

• Specificity testing: Using positive and negative control samples, including overexpression systems and knockout/knockdown cells when available.

• Cross-reactivity assessment: Particularly important when working with models from different species.

• Titration experiments: To determine optimal antibody concentration for each application.

• Epitope mapping: Understanding which region of the protein the antibody recognizes, especially important when studying different isoforms.

Since C17orf47 is synonymous with SEPTIN4 and has alternative names including ARTS and BRADEION, researchers should verify whether the antibody recognizes specific isoforms or all variants of the protein . Documentation from manufacturers regarding validation methods should be carefully reviewed before selecting an antibody for critical experiments.

What is the tissue expression profile of C17orf47/SEPTIN4?

C17orf47/SEPTIN4 is notably expressed in many human tissues, with significant expression reported in:

• Colon tissue

• Fallopian tube

• Various other tissues throughout the body

This broad tissue distribution makes C17orf47 antibodies valuable tools for comparative tissue studies. When planning immunohistochemistry experiments across multiple tissue types, researchers should consider the variable expression levels and optimize staining protocols accordingly. For quantitative comparisons between tissues, standardized protocols and careful normalization are essential to account for this differential expression.

What are the optimal parameters for using C17orf47 antibodies in immunohistochemistry?

For optimal results in immunohistochemistry applications with C17orf47 antibodies:

• Fixation method: Typically, 4% paraformaldehyde fixation is recommended, though some antibodies work well with formalin-fixed paraffin-embedded (FFPE) tissues.

• Antigen retrieval: Heat-induced epitope retrieval using citrate buffer (pH 6.0) is commonly effective.

• Antibody dilution: Starting dilutions of 1:100 to 1:500 are typically recommended, but optimal concentration should be determined experimentally for each application and tissue type .

• Detection system: Both chromogenic and fluorescent detection systems can be used depending on the research question.

• Controls: Include positive control tissues (colon, fallopian tube) and negative controls (primary antibody omission) .

The subcellular localization of C17orf47/SEPTIN4 (cytoplasmic vesicles and cytoplasm) should guide the interpretation of staining patterns . Non-specific nuclear staining should be considered with caution as it may represent background rather than true protein localization.

How can researchers optimize ELISA protocols using C17orf47 antibodies?

For developing robust ELISA assays using C17orf47 antibodies:

• Antibody selection: Consider using pairs of antibodies recognizing different epitopes for sandwich ELISA, or HRP/biotin-conjugated antibodies for direct detection .

• Coating concentration: Typically 1-10 μg/ml of capture antibody in carbonate buffer (pH 9.6).

• Blocking protocol: 3-5% BSA or non-fat milk in PBS to minimize background.

• Sample preparation: Cellular or tissue lysates should be prepared with appropriate protein extraction buffers containing protease inhibitors.

• Standard curve: Use recombinant C17orf47/SEPTIN4 protein for quantitative analysis.

• Detection optimization: For HRP-conjugated antibodies, determine the optimal substrate (TMB, ABTS) and development time .

The high purity (>95%) of commercially available antibodies helps ensure reliable ELISA results, but lot-to-lot variation should be monitored for long-term studies . Researchers should optimize dilutions based on their specific sample types and detection systems.

What are the considerations for using C17orf47 antibodies in apoptosis research?

Given C17orf47/SEPTIN4's role in apoptotic pathways, researchers should consider:

• Experimental timing: During apoptosis, protein localization and abundance may change rapidly; time-course experiments are recommended.

• Co-localization studies: Combine C17orf47 antibodies with markers of apoptotic machinery for comprehensive analysis.

• Isoform specificity: The eight identified isoforms may have different functions in apoptosis; verify which isoforms your antibody recognizes .

• Functional validation: Complement immunodetection with functional assays (caspase activation, TUNEL staining) to correlate protein detection with apoptotic events.

• Cell type considerations: Different cell types may exhibit variable expression and functions of C17orf47/SEPTIN4 during apoptosis.

Understanding that C17orf47/SEPTIN4 is also known as ARTS, a pro-apoptotic protein, provides context for its role in programmed cell death research . Carefully designed experimental controls are essential when investigating protein dynamics during the apoptotic process.

How should researchers approach troubleshooting when using C17orf47 antibodies?

When encountering challenges with C17orf47 antibodies, consider:

• Signal strength issues:

  • For weak signals: Increase antibody concentration, extend incubation time, enhance detection system, or try different antigen retrieval methods

  • For excessive background: Dilute antibody further, optimize blocking, reduce incubation time, or try alternative washing protocols

• Specificity concerns:

  • Verify antibody specificity using overexpression systems or knockdown controls

  • Consider pre-adsorption with the immunizing peptide if available

  • Test multiple antibodies targeting different epitopes

• Storage and handling:

  • Aliquot and store antibodies at -20°C, avoiding repeated freeze-thaw cycles

  • Protect HRP or fluorophore-conjugated antibodies from light exposure

  • Follow manufacturer recommendations for buffer composition and additives

• Sample preparation optimization:

  • Ensure complete protein denaturation for western blot applications

  • Optimize fixation protocols for immunohistochemistry/immunocytochemistry

  • Consider native versus denaturing conditions based on epitope accessibility

Maintaining detailed experimental records helps track variables that may affect antibody performance across different experiments and batches.

How can genetic variants in C17orf47/SEPTIN4 impact antibody-based research?

Genetic variants in C17orf47/SEPTIN4 can significantly impact antibody-based studies:

• Epitope alterations: Mutations may alter antibody binding sites, potentially resulting in false-negative results if the epitope is directly affected.

• Expression differences: Variants may affect protein expression levels, stability, or subcellular localization, altering detection patterns.

• Isoform variation: Genetic changes may influence alternative splicing, producing different isoform distributions that may not be detected by all antibodies .

Some variants in C17orf47 have been identified and analyzed in genetic studies, including those predicted to be benign by PolyPhen . When studying samples with potential genetic variation, researchers should ideally use antibodies targeting conserved regions or employ multiple antibodies recognizing different epitopes to ensure comprehensive detection.

What emerging techniques can enhance C17orf47 antibody-based research?

Researchers can leverage several advanced techniques to enhance C17orf47 antibody-based studies:

• Proximity ligation assays (PLA): For detecting protein-protein interactions involving C17orf47/SEPTIN4 with high specificity and sensitivity.

• Super-resolution microscopy: For detailed subcellular localization beyond the diffraction limit, particularly valuable for studying cytoplasmic vesicle associations.

• Mass cytometry (CyTOF): For multiparameter analysis of C17orf47/SEPTIN4 alongside numerous other markers in heterogeneous cell populations.

• Spatial transcriptomics combined with immunodetection: For correlating protein localization with gene expression patterns in tissue contexts.

• Antibody-based proteomics: For large-scale analysis of C17orf47/SEPTIN4 interactions and modifications.

These emerging approaches can provide deeper insights into the functional roles of C17orf47/SEPTIN4 in normal physiology and disease states beyond what traditional antibody applications can reveal.