CCT7 Antibody

Introduction to CCT7 Antibody

CCT7 antibodies are immunological reagents specifically designed to bind to and detect the CCT7 protein, a subunit of the chaperonin-containing TCP-1 (CCT) complex. These antibodies serve as valuable tools for researchers investigating protein folding mechanisms, cellular stress responses, and various disease processes. Available in both polyclonal and monoclonal formats, CCT7 antibodies enable precise detection of this important molecular chaperone in diverse experimental settings .

The development of high-quality CCT7 antibodies has facilitated significant advances in understanding the biological roles of this protein. These antibodies are produced using various immunization strategies, with synthetic peptides or recombinant fusion proteins serving as immunogens to generate specific immune responses against CCT7. This specificity allows researchers to study CCT7's expression, localization, and interactions in complex biological samples .

Functional Significance

The primary function of CCT7, as part of the TRiC complex, is to assist in the ATP-dependent folding of newly synthesized proteins. The complex forms a characteristic double-ring structure, creating a central cavity where unfolded polypeptides enter and undergo conformational changes to achieve their native, functional state . CCT7 plays a particularly important role in the folding of cytoskeletal proteins like actin and tubulin, which are essential for maintaining cellular structure and enabling intracellular transport .

Recent research has revealed that CCT7 is also involved in regulating the expression, maturation, and transport of G-protein coupled receptors (GPCRs) to the cell surface. Additionally, it prevents protein aggregation, a critical function for maintaining cellular homeostasis . The TRiC complex containing CCT7 also mediates the folding of WRAP53/TCAB1, thereby regulating telomere maintenance .

Polyclonal CCT7 Antibodies

Polyclonal CCT7 antibodies are produced by immunizing animals (typically rabbits) with CCT7-specific immunogens. These antibodies recognize multiple epitopes on the CCT7 protein, providing high sensitivity for detection applications. For example, Cepham Life Sciences offers a rabbit polyclonal antibody generated using a synthesized peptide derived from the internal region of human CCT7 . Similarly, Assay Genie's polyclonal antibody (CAB12146) is raised against a recombinant fusion protein corresponding to amino acids 310-500 of human CCT7 .

Monoclonal CCT7 Antibodies

Monoclonal CCT7 antibodies, such as Proteintech's 67540-1-Ig, are produced from a single B-cell clone, resulting in antibodies that recognize a single epitope on the CCT7 protein. This confers high specificity, though sometimes at the cost of reduced sensitivity compared to polyclonal alternatives. Monoclonal antibodies provide consistent results across experiments and are particularly valuable for applications requiring high reproducibility .

The production of monoclonal antibodies typically involves mouse immunization followed by hybridoma technology. These antibodies are extensively validated for specific applications such as Western blotting and immunofluorescence to ensure reliable performance .

Western Blotting

Western blotting represents one of the most common applications for CCT7 antibodies. This technique allows researchers to detect and quantify CCT7 protein in cell or tissue lysates. The recommended dilutions for CCT7 antibodies in Western blot applications vary by manufacturer, ranging from 1:500 to 1:50,000, depending on the specific antibody and sample type .

For instance, Proteintech's monoclonal CCT7 antibody (67540-1-Ig) has been validated for Western blotting at dilutions of 1:5,000 to 1:50,000 and successfully detects CCT7 in multiple cell lines including A549, NIH/3T3, HEK-293, LNCaP, HeLa, HepG, Jurkat, and K-562 cells . This wide reactivity profile makes it a versatile tool for comparative studies across different cellular models.

Immunofluorescence

CCT7 antibodies are effectively employed in immunofluorescence (IF) and immunocytochemistry (ICC) applications to visualize the subcellular localization of CCT7 protein. These techniques provide valuable insights into the spatial distribution of CCT7 within cells and tissues, helping researchers understand its functional contexts .

Research utilizing immunofluorescence with CCT7 antibodies has revealed that CCT7 primarily localizes in the cytoplasm, consistent with its role in cytosolic protein folding. When used in combination with antibodies against viral proteins like VP2 of canine parvovirus, IF has demonstrated co-localization between CCT7 and these viral components, suggesting functional interactions .

Co-Immunoprecipitation

Co-immunoprecipitation (Co-IP) experiments using CCT7 antibodies have been instrumental in identifying protein-protein interactions involving CCT7. This application has revealed important biological relationships, such as the interaction between CCT7 and the VP2 protein of canine parvovirus .

In a study investigating viral replication mechanisms, researchers used CCT7 antibodies to perform Co-IP experiments that confirmed the physical interaction between CCT7 and viral VP2 proteins. This technique helped establish the role of CCT7 in viral replication processes and identified the specific region of VP2 (amino acids 231-320) that interacts with CCT7 .

ELISA

Enzyme-linked immunosorbent assay (ELISA) represents another important application for CCT7 antibodies. This technique allows for quantitative measurement of CCT7 protein levels in biological samples, providing a high-throughput approach for screening multiple samples .

Immunogen Information

The immunogens used to generate CCT7 antibodies vary across manufacturers, influencing the specific epitopes recognized. For example, Cepham Life Sciences' polyclonal antibody is raised against a synthesized peptide derived from the internal region of human CCT7 . Assay Genie's polyclonal antibody (CAB12146) uses a recombinant fusion protein corresponding to amino acids 310-500 of human CCT7 . Proteintech's monoclonal antibody is produced using a TCP1 eta fusion protein (Ag9502) .

Species Reactivity and Cross-Reactivity

The species reactivity of CCT7 antibodies is an important consideration when selecting an appropriate reagent for experimental applications. Table 1 summarizes the documented reactivity of various commercial CCT7 antibodies:

This cross-reactivity information is crucial for researchers working with different model systems, as it indicates which antibodies are suitable for detecting CCT7 in specific species .

CCT7 in Hepatocellular Carcinoma

Research utilizing CCT7 antibodies has revealed significant associations between CCT7 expression and hepatocellular carcinoma (HCC). Multiple studies have demonstrated that CCT7 is consistently upregulated in HCC tissues compared to normal liver tissues .

A comprehensive analysis of CCT7 expression in HCC revealed that CCT7 mRNA levels were significantly higher in tumor tissues than in adjacent normal tissues. This finding was validated at the protein level using immunohistochemistry with CCT7 antibodies. The study documented a 1.63-fold increase in CCT7 expression in HCC samples compared to normal liver tissues (P=9.00E-41) .

CCT7 in Viral Replication

CCT7 antibodies have been instrumental in elucidating the role of CCT7 in viral replication processes. Research on canine parvovirus (CPV) has demonstrated that CCT7 interacts with the viral VP2 protein, playing a crucial role in viral replication .

Co-immunoprecipitation experiments using CCT7 antibodies confirmed the physical interaction between CCT7 and VP2. Further investigations revealed that CCT7 enhances the stability of VP2 protein, as demonstrated by cycloheximide chase experiments. When CCT7 expression was knocked down using RNA interference or inhibited by HSF1A, VP2 protein levels decreased significantly. Conversely, overexpression of CCT7 led to increased VP2 expression .

This research highlights a potential mechanism by which host cell factors facilitate viral replication and suggests that CCT7 could represent a therapeutic target for preventing and controlling viral infections like CPV.

Other Disease Associations

In addition to HCC and viral infections, CCT7 has been linked to several other pathological conditions. GeneCards data indicates associations between CCT7 and myocardial infarction and babesiosis . These connections suggest broader implications for CCT7 in various disease processes, although the precise mechanisms remain to be fully elucidated.

CCT7 as a Diagnostic Marker

Recent research has explored the potential of CCT7 as a diagnostic marker, particularly for HCC. In a study examining the diagnostic value of CCT7 expression, researchers found that CCT7 could identify HCC with greater accuracy than traditional markers like alpha-fetoprotein (AFP), especially in early-stage disease and in patients with low AFP expression .

Receiver operating characteristic curve analyses indicated that CCT7 expression had better diagnostic value than AFP for early-stage HCC patients and those with low AFP expression. The positive predictive value of CCT7 expression was higher than that of AFP expression, suggesting its potential utility as a complementary or alternative diagnostic marker .

CCT7 in Protein Folding Pathways

Research using CCT7 antibodies has contributed to our understanding of protein folding pathways and their dysregulation in disease. As a component of the TRiC complex, CCT7 participates in the folding of approximately 10% of newly synthesized proteins, including many that are essential for cellular function .

Studies have revealed that CCT7, along with other CCT subunits, is frequently upregulated in cancer tissues. For instance, analysis of multiple datasets showed consistent elevation of CCT7 in HCC compared to normal liver tissues across different cohorts. In the Roessler Liver 2 dataset, CCT7 was found to be upregulated in HCC tissues with a fold change of 1.63 (P=9.00E-41) .