CCDC24 Antibody

What is CCDC24 protein and why is it of interest to researchers?

CCDC24, or coiled-coil domain-containing protein 24, is involved in various cellular processes including cell proliferation, differentiation, cell cycle regulation, and DNA damage response. Its potential role in cancer development and progression makes it a significant target for research into cancer biology and therapeutic development. The protein's involvement in these fundamental cellular processes provides researchers with opportunities to understand critical regulatory mechanisms in both normal and pathological contexts .

What types of CCDC24 antibodies are available for research applications?

Several types of CCDC24 antibodies are available for research use. These include polyclonal antibodies such as the PACO38534 from Assay Genie and AP18327a from Abcepta. These antibodies are typically raised in rabbits against human CCDC24 protein. The antibodies may target different regions of the protein, with some specific to the N-terminal region (amino acids 1-30), while others recognize the full-length protein (1-307 amino acids). These are available in unconjugated forms as well as conjugated to various labels including FITC, Biotin, APC, and HRP depending on experimental requirements .

What experimental applications are CCDC24 antibodies validated for?

CCDC24 antibodies have been validated for several experimental applications including:

• Western Blotting (WB): For detecting CCDC24 protein in cell or tissue lysates

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

• IHC (Immunohistochemistry): For localization of the protein in tissue sections

• E (ELISA): For specific detection in complex samples

The recommended dilutions vary by application, typically ranging from 1:500-1:2000 for Western blot, 1:2000-1:10000 for ELISA, and 1:20-1:200 for IHC, though researchers should optimize these for their specific experimental conditions .

How should I design experiments to validate CCDC24 antibody specificity for my research?

To validate CCDC24 antibody specificity, implement a multi-step approach:

• Positive and negative controls: Use cell lines known to express CCDC24 (e.g., MCF-7 has been documented) as positive controls and include knockout/knockdown samples as negative controls.

• Epitope competition assay: Pre-incubate the antibody with the immunizing peptide before application to demonstrate binding specificity.

• Multiple antibody validation: Compare results using antibodies targeting different epitopes of CCDC24 (such as N-terminal specific vs. full-length protein antibodies).

• Western blot analysis: Confirm single-band specificity at the expected molecular weight (calculated MW is approximately 34kDa).

• Cross-reactivity assessment: Test the antibody on samples from other species if cross-reactivity is claimed by the manufacturer .

What are the optimal storage and handling conditions for maintaining CCDC24 antibody functionality?

For optimal CCDC24 antibody preservation and functionality:

• Short-term storage (up to 2 weeks): Maintain refrigerated at 2-8°C.

• Long-term storage: Store at -20°C in small aliquots to prevent repeated freeze-thaw cycles which can damage the antibody.

• Buffer conditions: Most CCDC24 antibodies are supplied in PBS with 0.09% sodium azide and may contain glycerol (up to 50%) as a cryoprotectant.

• Handling: Avoid repeated freeze-thaw cycles by preparing single-use aliquots immediately upon receipt.

• Working dilutions: Prepare fresh working dilutions on the day of the experiment for optimal results.

• Temperature transitions: Allow the antibody to equilibrate to room temperature before opening to prevent condensation .

What are the recommended protocols for using CCDC24 antibodies in Western blot applications?

For optimal Western blot results with CCDC24 antibodies:

• Sample preparation:

  • Lyse cells in RIPA buffer with protease inhibitors

  • Determine protein concentration (BCA/Bradford assay)

  • Load 20-50μg total protein per lane

• Gel electrophoresis:

  • Use 10-12% polyacrylamide gels

  • Include positive control (e.g., MCF-7 cell lysate)

• Transfer and blocking:

  • Transfer to PVDF membrane (preferred over nitrocellulose)

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

• Primary antibody incubation:

  • Dilute antibody 1:500-1:2000 in blocking buffer

  • Incubate overnight at 4°C with gentle agitation

• Detection:

  • Use appropriate HRP-conjugated secondary antibody (anti-rabbit IgG)

  • Visualize using enhanced chemiluminescence

• Expected result:

  • CCDC24 should appear as a single band at approximately 34kDa .

How can CCDC24 antibodies be employed in studies examining the relationship between CCDC24 and cancer progression?

For investigating CCDC24's role in cancer progression:

• Expression profiling across cancer stages:

  • Use IHC with CCDC24 antibodies on tissue microarrays containing samples from different cancer stages

  • Quantify expression levels using digital pathology software

  • Correlate expression with clinical parameters including tumor stage, grade, and patient survival

• Mechanistic studies:

  • Combine CCDC24 immunodetection with markers of cell cycle (Ki67, cyclins) and DNA damage response (γH2AX, 53BP1)

  • Implement dual immunofluorescence to assess co-localization with other proteins

  • Use proximity ligation assays to detect protein-protein interactions involving CCDC24

• Response to treatment:

  • Monitor CCDC24 expression before and after treatment with chemotherapeutic agents

  • Correlate changes in expression with treatment response

• Functional validation:

  • Combine antibody-based detection with gene expression modulation (overexpression/knockdown)

  • Assess changes in cellular phenotypes including proliferation, migration, and apoptosis .

What methodological considerations are important when using CCDC24 antibodies in immunohistochemistry applications?

Critical methodological considerations for IHC with CCDC24 antibodies:

• Tissue preparation and fixation:

  • Optimal fixation: 10% neutral buffered formalin for 24-48 hours

  • Paraffin embedding with standard protocols

  • Section thickness: 4-5μm for optimal staining

• Antigen retrieval methods:

  • Heat-induced epitope retrieval (HIER) using citrate buffer (pH 6.0) or EDTA buffer (pH 9.0)

  • Pressure cooker treatment for 15-20 minutes recommended

• Blocking strategies:

  • Block endogenous peroxidase with 3% H₂O₂

  • Use species-appropriate serum (5-10%) to reduce background

  • Include avidin/biotin blocking if using biotin-based detection systems

• Antibody optimization:

  • Test multiple dilutions (starting range 1:20-1:200)

  • Optimize incubation time and temperature (overnight at 4°C or 1-2 hours at room temperature)

• Detection systems:

  • Polymer-based detection systems provide better signal-to-noise ratio than ABC methods

  • Consider tyramide signal amplification for low-abundance proteins

• Controls:

  • Include positive tissue controls with known CCDC24 expression

  • Use isotype control antibodies at the same concentration

  • Include negative controls omitting primary antibody .

How can researchers utilize CCDC24 antibodies in conjunction with Mendelian Randomization approaches to understand disease mechanisms?

Integrating CCDC24 antibodies with Mendelian Randomization approaches:

• Genotype-protein expression correlation:

  • Use CCDC24 antibodies to quantify protein levels in samples with known SNP genotypes

  • Correlate specific SNPs (identified in GWAS studies) with CCDC24 protein expression

  • Establish whether genetic variants influence protein abundance or post-translational modifications

• Tissue-specific expression analysis:

  • Apply IHC or Western blot in multiple tissue types from individuals with different genotypes

  • Determine whether genetic associations with disease are mediated through tissue-specific expression patterns

• Functional validation of causal SNPs:

  • Employ CCDC24 antibodies to assess how manipulation of causal SNPs affects protein expression

  • Use CRISPR-Cas9 gene editing to introduce SNP variants and monitor changes in protein expression

• Protein-protein interaction studies:

  • Implement co-immunoprecipitation with CCDC24 antibodies to identify interaction partners

  • Compare interaction profiles between different genotypes to identify mechanism-based differences

• Longitudinal studies:

  • Measure CCDC24 protein levels at multiple timepoints to establish temporal relationships

  • Combine with genetic instruments to strengthen causal inference .

What are common issues encountered when using CCDC24 antibodies and how can they be resolved?

Common issues and their solutions when working with CCDC24 antibodies:

These troubleshooting approaches should be systematically implemented while maintaining appropriate controls throughout .

How should researchers interpret varying CCDC24 antibody staining patterns across different cell types or tissues?

Interpreting differential CCDC24 staining patterns requires careful consideration:

• Biological relevance assessment:

  • Cell type-specific expression patterns may reflect tissue-specific functions

  • Subcellular localization differences (nuclear vs. cytoplasmic) may indicate different functional states

  • Expression intensity correlations with developmental stages or pathological conditions provide functional insights

• Technical validation:

  • Confirm pattern specificity using multiple antibodies targeting different epitopes

  • Complement antibody-based detection with mRNA expression analysis (RT-PCR, RNA-seq)

  • Validate unusual patterns with orthogonal techniques (e.g., GFP-tagged protein expression)

• Physiological context interpretation:

  • Cell cycle dependency: Synchronize cells and assess expression at different cell cycle phases

  • Stress response: Compare patterns before and after cellular stress (oxidative, genotoxic)

  • Differentiation status: Correlate with markers of cellular differentiation

• Pathological significance:

  • Compare normal vs. diseased tissues systematically

  • Quantify pattern changes using digital image analysis

  • Correlate with clinical parameters for potential biomarker applications .

How can researchers differentiate between specific and non-specific binding when using CCDC24 antibodies?

To distinguish specific from non-specific binding with CCDC24 antibodies:

• Peptide competition assays:

  • Pre-incubate the antibody with excess immunizing peptide

  • Specific binding should be significantly reduced or eliminated

  • Non-specific binding will likely remain unchanged

• Knockout/knockdown validation:

  • Use CRISPR/Cas9 knockout or siRNA knockdown of CCDC24

  • Compare staining patterns between wild-type and knockout/knockdown samples

  • Specific signals should be substantially reduced in knockout/knockdown samples

• Multiple antibody confirmation:

  • Use antibodies targeting different epitopes of CCDC24

  • Specific binding patterns should be consistent across antibodies

  • Discrepancies may indicate non-specific binding or post-translational modifications

• Signal correlation with expression level:

  • Compare antibody signal intensity with known CCDC24 expression levels across different cell lines

  • Positive correlation supports specificity

  • Random pattern suggests non-specific binding

• Isotype controls:

  • Use matched isotype control antibodies at the same concentration

  • Helps identify Fc-receptor mediated non-specific binding

  • Particularly important for flow cytometry and IHC applications .

How can CCDC24 antibodies be utilized to investigate its potential role in cervical carcinoma pathogenesis?

For investigating CCDC24's role in cervical carcinoma:

• Expression profiling in disease progression:

  • Perform IHC analysis on tissue microarrays containing normal cervical tissue, cervical intraepithelial neoplasia (CIN) of various grades, and invasive carcinoma

  • Quantify expression levels and correlate with disease stage

  • Assess co-expression with HPV markers (E6/E7) to investigate virus-host interactions

• Mechanistic studies:

  • Combine CCDC24 antibody detection with antibodies against DNA damage response proteins (γH2AX, BRCA1, 53BP1)

  • Investigate potential interactions between CCDC24 and immune response pathways using dual immunofluorescence

  • Explore CCDC24's relationship with antibody-mediated immune responses in cervical carcinoma patients

• Genetic correlation studies:

  • Stratify tissue samples based on SNPs identified in Mendelian Randomization studies

  • Compare CCDC24 protein expression levels between different genotypes

  • Investigate whether specific SNPs (e.g., GCST90006901, GCST90006909) affect protein expression or localization

• Response to therapy:

  • Monitor CCDC24 expression before and after standard treatments

  • Evaluate potential as a predictive biomarker for treatment response

  • Explore relationships between CCDC24 expression and immune checkpoint markers .

What experimental approaches can be used to investigate the relationship between CCDC24 and antibody-mediated immune responses?

To investigate CCDC24's relationship with antibody-mediated immunity:

• Co-expression analysis:

  • Use multi-color immunofluorescence to assess CCDC24 expression in immune cell populations

  • Focus on B cells, plasma cells, and follicular dendritic cells

  • Correlate CCDC24 expression with markers of B cell activation and antibody production

• In vitro functional assays:

  • Modulate CCDC24 expression in B cell lines using overexpression/knockdown approaches

  • Assess effects on antibody production, class switching, and somatic hypermutation

  • Measure changes in signaling pathways involved in B cell activation

• Patient-derived samples:

  • Compare CCDC24 expression in peripheral blood B cells from healthy donors versus patients with autoimmune diseases

  • Correlate expression levels with serum antibody titers and autoantibody presence

  • Investigate genetic associations between CCDC24 SNPs and antibody-mediated conditions

• Animal models:

  • Generate CCDC24 knockout or transgenic mouse models

  • Characterize B cell development, antibody responses to antigens, and susceptibility to antibody-mediated diseases

  • Perform adoptive transfer experiments to isolate B cell-intrinsic effects

• Mendelian Randomization approach:

  • Utilize bidirectional MR to further elucidate causal relationships

  • Focus on SNPs with established associations (GCST90006891, GCST90006894, GCST90006899, GCST90006911)

  • Integrate protein expression data with genetic findings .

What considerations are important when using CCDC24 antibodies in studying potential cancer biomarkers?

Critical considerations for CCDC24 as a cancer biomarker:

• Analytical validation:

  • Establish rigorous validation protocols using multiple CCDC24 antibodies

  • Determine sensitivity, specificity, reproducibility, and robustness across different sample types

  • Standardize scoring methods for quantitative assessment

• Clinical validation:

  • Perform retrospective analysis on well-annotated clinical cohorts

  • Include samples from diverse patient populations and disease stages

  • Correlate expression with established prognostic factors and clinical outcomes

• Technical standardization:

  • Develop standard operating procedures for sample preparation and staining

  • Establish quality control metrics for antibody lot-to-lot variation

  • Implement automated image analysis to reduce subjective interpretation

• Biological context:

  • Consider the impact of tumor heterogeneity on expression patterns

  • Assess expression in the context of tumor microenvironment

  • Evaluate stability of expression over disease course and treatment

• Comparison with existing biomarkers:

  • Perform head-to-head comparisons with established biomarkers

  • Assess incremental value when added to existing prognostic models

  • Evaluate cost-effectiveness and clinical utility

• Potential for targeted therapy:

  • Investigate whether CCDC24 expression levels predict response to specific treatments

  • Explore CCDC24 as a potential therapeutic target itself

  • Develop companion diagnostic approaches if therapeutic relevance is established .