CCDC127 Antibody

What is CCDC127 and why is it studied?

CCDC127 (Coiled-coil domain-containing protein 127) is a protein encoded by the CCDC127 gene in humans. The canonical protein has a reported length of 260 amino acid residues and a molecular mass of approximately 30.8 kDa. It is notable for its wide expression pattern across many tissue types, making it relevant for diverse research areas . While the specific function of CCDC127 remains under investigation, researchers use antibodies against this protein to study its expression patterns, localization, and potential role in cellular processes.

What are the key identifiers for the human CCDC127 protein?

Human CCDC127 protein can be identified through several database identifiers:

• UniProt ID: Q96BQ5

• UniProt Entry Name: CC127_HUMAN

• Entrez Gene ID: 133957

• KEGG: hsa:133957

• String: 9606.ENSP00000296824

The protein is also known by synonyms including "coiled-coil domain-containing protein 127" and "FLJ25701" .

How conserved is CCDC127 across species?

CCDC127 shows significant conservation across multiple species, making cross-species studies possible. Based on sequence identity, the human CCDC127 has:

• 83% sequence identity with mouse ortholog

• 82% sequence identity with rat ortholog

• 86% sequence identity with bovine ortholog

Additionally, CCDC127 gene orthologs have been reported in frog, chimpanzee, and chicken species , indicating evolutionary conservation across diverse vertebrates.

What are the primary research applications for CCDC127 antibodies?

CCDC127 antibodies are primarily used in:

• Western Blot (WB): For detecting CCDC127 protein in lysates and determining expression levels

• Immunohistochemistry (IHC): For examining tissue localization patterns

• Immunohistochemistry-Paraffin (IHC-P): For detecting the protein in fixed, paraffin-embedded tissues

• Immunocytochemistry (ICC): For examining subcellular localization

• Enzyme-Linked Immunosorbent Assay (ELISA): For quantitative protein detection

• Immunofluorescence: For visualizing protein distribution within cells

Each application provides unique insights into CCDC127 expression, localization, and potential functions in different experimental contexts.

What are the recommended antibody dilutions for different applications?

Based on validated antibody products, the following dilution ranges are recommended:

These ranges provide starting points, but optimal dilutions should be determined experimentally for each specific antibody and application .

How should researchers approach CCDC127 antibody validation?

For rigorous validation of CCDC127 antibodies, researchers should:

• Confirm specificity using positive and negative controls

• Verify the expected molecular weight (approximately 30.8-31 kDa) in Western blot experiments

• Test cross-reactivity if working with non-human samples

• Validate across multiple applications when possible

• Consider using genetically modified systems (knockouts or overexpression) for definitive validation

• Document all validation parameters including antibody concentration, incubation conditions, and detection methods

Proper validation ensures reliable and reproducible results in subsequent experiments.

What are the optimal storage conditions for CCDC127 antibodies?

For maximum stability and activity, CCDC127 antibodies should be:

• Aliquoted upon receipt to avoid repeated freeze-thaw cycles

• Stored at -20°C or -80°C for long-term storage

• Protected from light if conjugated to fluorophores (like FITC)

• Kept in appropriate buffer conditions, typically including glycerol as a cryoprotectant

Most commercial CCDC127 antibodies are supplied in buffers containing glycerol (typically 20-50%) and preservatives like thimerosal or Proclin-300 to maintain stability during storage .

What controls should be included when using CCDC127 antibodies?

For rigorous research, the following controls should be included:

• Positive control: Tissue or cell line known to express CCDC127 (the protein is widely expressed across many tissues)

• Negative control:

  • Primary antibody omission control

  • Isotype control (matching the host species and isotype of the CCDC127 antibody)

  • Blocking peptide control when available

• Loading control: For Western blot normalization

• Specificity control: Pre-absorption with immunizing peptide when available

These controls help validate results and address potential non-specific binding or background issues.

How can researchers select the appropriate CCDC127 antibody format for their experiment?

Selection should be based on:

• Application requirements:

  • Unconjugated antibodies: Suitable for Western blot, IHC, and applications requiring secondary detection

  • Conjugated antibodies (e.g., FITC-conjugated): Ideal for direct immunofluorescence without secondary antibodies

• Detection system:

  • For fluorescence applications: Consider excitation/emission properties (FITC-conjugated CCDC127 antibodies have excitation/emission at 499/515nm)

  • For enzymatic detection: Unconjugated antibodies with appropriate secondaries

• Host species compatibility:

  • Available CCDC127 antibodies are predominantly rabbit-derived

  • Consider species compatibility with other antibodies in multi-labeling experiments

How can CCDC127 antibodies be integrated into multi-parameter analyses?

For comprehensive studies, researchers can:

• Combine CCDC127 detection with other markers in multi-color immunofluorescence:

  • FITC-conjugated CCDC127 antibody (excitation/emission: 499/515nm, laser line: 488nm) can be paired with spectrally distinct fluorophores

  • Consider antibody host species to avoid cross-reactivity in multi-labeling experiments

• Incorporate into proteomics workflows:

  • Use for immunoprecipitation followed by mass spectrometry to identify interaction partners

  • Combine with subcellular fractionation to determine precise localization

• Implement in high-content imaging:

  • Quantify expression levels across different experimental conditions

  • Analyze colocalization with other proteins of interest

What approaches can be used to study CCDC127 in the context of chromatin organization?

While CCDC127's specific role in chromatin organization is not directly established in the search results, researchers interested in this direction could:

• Investigate potential relationships between CCDC127 and chromatin regulators like cohesin

  • Cohesin complexes play crucial roles in chromatin loop formation and gene regulation

  • Techniques like chromatin immunoprecipitation (ChIP) could reveal potential associations

• Examine CCDC127 localization relative to nuclear architecture:

  • Super-resolution microscopy with CCDC127 antibodies could reveal spatial relationships to chromatin domains

  • Co-immunoprecipitation experiments might identify interactions with chromatin-associated proteins

• Study CCDC127 in the context of V(D)J recombination:

  • V(D)J recombination depends on chromatin configuration for generating antibody diversity

  • Investigating potential roles of CCDC127 in immune cells could be particularly relevant

How can genetic approaches complement CCDC127 antibody studies?

Integrating genetic approaches with antibody-based detection can provide more comprehensive insights:

• CRISPR/Cas9-mediated gene editing:

  • Generate CCDC127 knockout or tagged cell lines

  • Validate antibody specificity using knockout controls

  • Study phenotypic consequences of CCDC127 depletion or modification

• Overexpression systems:

  • Express tagged versions (e.g., GFP-CCDC127) for live-cell imaging

  • Compare antibody staining patterns with tagged protein localization

  • Study effects of CCDC127 upregulation on cellular processes

• RNA interference:

  • Use siRNA or shRNA to knockdown CCDC127

  • Confirm knockdown efficiency by antibody-based detection

  • Investigate functional consequences of reduced CCDC127 expression

What are common challenges when using CCDC127 antibodies and how can they be addressed?

Researchers may encounter several technical challenges:

• Background or non-specific staining:

  • Optimize blocking conditions (typically 5% BSA or normal serum)

  • Titrate antibody concentration (start with manufacturer recommendations like 1:500-1:3000 for WB)

  • Include appropriate controls (isotype control, secondary-only control)

  • Consider using alternative detection systems

• Weak or absent signal:

  • Increase antibody concentration within recommended ranges

  • Extend incubation time (overnight at 4°C often improves sensitivity)

  • Optimize antigen retrieval for IHC-P applications

  • Consider more sensitive detection systems

• Unexpected band size in Western blot:

  • Verify sample preparation and protein denaturation conditions

  • Consider post-translational modifications that may affect migration

  • The expected molecular weight for CCDC127 is approximately 31 kDa

How should researchers approach conflicting results with different CCDC127 antibodies?

When different antibodies yield conflicting results:

• Compare antibody epitopes:

  • Some antibodies target N-terminal regions while others target central regions

  • Different epitopes may be differentially accessible in various experimental conditions

• Validate with orthogonal approaches:

  • Complement antibody studies with RNA-level analysis (qPCR, RNA-seq)

  • Use tagged expression constructs as independent confirmation

  • Apply genetic approaches (CRISPR/Cas9 knockouts) to confirm specificity

• Systematically document conditions:

  • Record fixation methods, buffer compositions, and incubation parameters

  • Test multiple antibodies under identical conditions

  • Consider context-dependent protein modifications or interactions