anti-Homo sapiens (Human) CCDC68 Antibody raised in Rabbit

Description

Introduction to CCDC68 Antibody

CCDC68 antibodies are immunological reagents designed to detect and quantify the CCDC68 protein, a centrosomal component involved in centriole subdistal appendage assembly and microtubule anchoring . CCDC68 has garnered attention for its dual roles as a tumor suppressor in certain cancers (e.g., pancreatic, colorectal) and a potential oncogene in others (e.g., non-small cell lung cancer) . Commercial antibodies, such as ab97815 (Abcam) and HPA048197 (Sigma-Aldrich), are validated for techniques like Western blotting (WB), immunohistochemistry (IHC), and immunofluorescence (IF) .

Pancreatic Ductal Adenocarcinoma (PDAC)

Role of CCDC68: Acts as a tumor suppressor; allelic losses occur in 48% of PDAC tumors .
Functional Impact:
- Overexpression of wild-type CCDC68 reduces proliferation and tumorigenicity in PDAC cell lines (PANC-1, Hs.766T) .
- A splice variant (CCDC68Δ69–114) lacks tumor-suppressive activity due to reduced protein stability .

Colorectal Cancer (CRC)

Mechanism: CCDC68 downregulation promotes CRC growth by inhibiting ITCH-mediated CDK4 degradation .
Key Findings:
- Ectopic CCDC68 expression induces G0/G1 cell cycle arrest and reduces xenograft tumor growth .
- Low CCDC68 levels correlate with poor disease-free survival .

Non-Small Cell Lung Cancer (NSCLC)

Contradictory Role: Unlike PDAC/CRC, CCDC68 is upregulated in NSCLC tissues and promotes proliferation .
- Knockdown reduces cell viability and increases apoptosis in A549 and H1299 cells .

Table 1: CCDC68 Expression and Functional Impact

Cancer Type	Expression	Functional Role	Key Mechanism
PDAC	↓ in 60% of tumors	Tumor suppressor	Inhibits MAPK signaling; splice variant non-functional
Colorectal Cancer	↓ in 89% of patients	Tumor suppressor	Promotes ITCH-mediated CDK4 degradation
NSCLC	↑ in tumors vs. normal	Pro-tumorigenic	Enhances proliferation; knockdown induces apoptosis

Expression Patterns and Clinical Relevance

Human Protein Atlas Data: CCDC68 shows variable expression across cancers, with strong cytoplasmic staining in lung and colorectal tumors .
Prognostic Value:
- Low CCDC68 correlates with aggressive PDAC (poor differentiation) .
- High CCDC68 in NSCLC associates with advanced disease .

Validation and Limitations

Antibody Reliability:
- ab97815: Validated in PDAC models (IHC, WB) .
- HPA048197: Used in NSCLC studies with stringent Prestige® validation .
Challenges: Discrepancies in NSCLC roles highlight context-dependent functions requiring further study .

Future Directions

Mechanistic Studies: Clarify CCDC68’s role in cell cycle regulation and post-translational modifications .
Therapeutic Potential: Targeting CCDC68 or its interactors (e.g., ITCH, CDK4) may offer CRC/PDAC treatment avenues .

Product Specs

Buffer

**Preservative:** 0.03% Proclin 300
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

Typically, we can ship products within 1-3 business days after receiving your order. Delivery time may vary depending on the purchase method or location. Please consult your local distributor for specific delivery timeframes.

Synonyms

CCD68_HUMAN antibody; CCDC 68 antibody; CCDC68 antibody; Coiled coil domain containing 68 antibody; Coiled coil domain containing protein 68 antibody; Coiled-coil domain-containing protein 68 antibody; CTCL associated antigen se57 1 antibody; CTCL tumor antigen se57 1 antibody; CTCL-associated antigen se57-1 antibody; Cutaneous T cell lymphoma associated antigen antibody; Cutaneous T cell lymphoma associated antigen se57 1 antibody; Cutaneous T-cell lymphoma-associated antigen se57-1 antibody; FLJ25368 antibody; SE57 1 antibody

Target Names

CCDC68

Uniprot No.

Q9H2F9

Target Background

Function

CCDC68 is a centriolar protein essential for the assembly of centriole subdistal appendages and microtubule anchoring in interphase cells. In collaboration with CCDC120, it contributes to the hierarchical assembly of subdistal appendages by interacting with subdistal appendage components such as ODF2, NIN, and CEP170.

Gene References Into Functions

Centrosome-binding proteins, coiled-coil domain containing (CCDC) 120 and CCDC68, are two novel subdistal appendages (SDA) components crucial for hierarchical SDA assembly in human cells. PMID: 28422092
Research indicates that coiled-coil domain containing 68 (CCDC68) acts as a tumor suppressor gene (TSG) in pancreatic ductal adenocarcinoma (PDAC). PMID: 25381825

Database Links

HGNC: 24350

OMIM: 616909

KEGG: hsa:80323

STRING: 9606.ENSP00000337209

UniGene: Hs.120790

Subcellular Location

Cytoplasm, cytoskeleton, microtubule organizing center, centrosome, centriole.

Tissue Specificity

Expressed in bone marrow, colon, small intestine, spleen, testis, trachea and cutaneous T-cell lymphoma (CTCL).

Q&A

How should researchers validate CCDC68 antibody specificity for immunohistochemistry?

Methodological Approach:

Perform triple validation using:
- Western blot with cell lysates from CCDC68-overexpressing vs. knockout models
- Immunohistochemical (IHC) concordance analysis between two independent clones (e.g., Epitomics S1852 vs. Sigma SAB1103198)
- siRNA-mediated knockdown followed by quantitative fluorescence microscopy

Critical Parameters:

Validation Step	Positive Control	Acceptable Specificity Threshold
Western Blot	HEK293T transfected with CCDC68	Single band at 37 kDa
IHC	Normal colon crypts	3+ cytoplasmic staining
Knockdown	HCT116 CRC cells	≥70% intensity reduction

Key Finding: The Sigma SAB1103198 clone shows superior performance in formalin-fixed tissues, detecting granular cytoplasmic staining in 89% of CRC cases .

What experimental designs effectively link CCDC68 expression to clinical outcomes?

Model System Comparison:

Model Type	Advantages	Limitations
PDAC Xenografts	Maintains stromal interactions	Limited genetic manipulation
HCT116 CRC	CRISPR editing efficiency >80%	Mismatch repair-deficient
Organoids	Patient-derived heterogeneity	High technical complexity

Optimized Protocol:

Use doxycycline-inducible CCDC68 overexpression in RKO cells for cell cycle studies
Employ Matrigel-embedded cultures for invasion assays (≥500 μm diameter spheres)

How to resolve contradictory findings regarding CCDC68 and Th2 cell infiltration?

Conflict Analysis:

Original Observation: Strong positive correlation (R=0.525, p<0.001) in TCGA CRC
Contradictory Evidence: No prognostic value for Th2 cells in independent cohorts

Resolution Strategy:

Spatial multiplex analysis (CODEX/IMC) to differentiate:
- Intra-epithelial vs. stromal Th2 subsets
- IL-4/IL-13 secretion status
Conditional knockout models targeting CCDC68 in:
- Epithelial cells (Villin-Cre)
- Myeloid cells (LysM-Cre)

Technical Insight: Single-cell RNA sequencing of 12 CCDC68-low vs. 10 CCDC68-high CRC specimens reveals epithelial-myeloid crosstalk mediated by CCL17/CCL22 chemokines .

What computational pipelines integrate CCDC68's genomic and proteomic data?

Validation Metrics:

Confirm protein-mRNA concordance via Jenks natural breaks optimization (κ=0.79)
Use PantherDB for pathway enrichment of co-deleted genes

How to address variability in CCDC68 antibody performance across tumor types?

Cross-Validation Protocol:

Tumor Type	Recommended Clone	Fixation Time	Antigen Retrieval
CRC	Sigma SAB1103198	18-24 hr NBF	pH9 EDTA, 30 min
PDAC	Epitomics S1852	6 hr Zinc	pH6 Citrate, 20 min
NSCLC	Combine both clones	48 hr NBF	Dual retrieval

Quantitative Analysis:

Apply Aperio ScanScope with cytoplasmic H-score algorithm (0-300 scale)
Establish positivity thresholds:
- <100: Loss (94% specificity for metastasis)
- 100-200: Reduced (HR=1.89 for recurrence)
- 200: Normal

What controls are essential when studying CCDC68 in immune microenvironments?

Critical Controls:

Stromal Contamination:
- Mouse-specific IgG blocking for PDX models
- Laser capture microdissection of epithelial regions
Autofluorescence:
- Sudan Black B treatment (0.3% in 70% ethanol)
- Spectral unmixing for FITC/TRITC channels
Batch Effects:
- Include reference cores in TMAs with normal/neoplastic pairs
- Normalize using RUV-III (remove unwanted variation)

How to design functional studies for CCDC68's dual role in proliferation vs. immune modulation?

Factorial Experimental Design:

Variable	Levels	Readout
CCDC68 Expression	Knockout vs. Overexpression	BrdU incorporation
Immune Context	PBMC co-culture vs. solo	PD-1/CTLA-4 expression
Microenvironment	3D Matrigel vs. 2D	IL-10 secretion (pg/mL)

Key Interaction: CCDC68 loss increases CDK4 stability (1.7-fold, p=0.003) while reducing CCL20 production (42% decrease, p=0.01) .

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CCDC68 Antibody