Tumor Protein p53 (p53) Panel Screening Service

Genetic testing forms the bedrock of our p53 screening, employing high-resolution DNA sequencing to meticulously analyze the TP53 gene.

• Germline Testing: This is indispensable for diagnosing inherited cancer predisposition syndromes. By analyzing DNA derived from non-tumor samples, such as blood or saliva, we can identify constitutional TP53 mutations that confer a significantly elevated lifetime risk of developing various cancers. This information is crucial for genetic counseling and proactive surveillance strategies.
• Somatic Testing: For established malignancies, somatic testing involves the analysis of DNA extracted directly from tumor tissue. This identifies acquired TP53 mutations that drive tumor development and progression. The presence and specific nature of these somatic mutations can serve as vital prognostic markers, influencing disease aggressiveness and predicting response to specific chemotherapeutic agents or targeted therapies.

Beyond mere mutation detection, understanding the functional consequence of TP53 alterations is paramount. Creative Biolabs employs a suite of functional assays to evaluate the transcriptional activity and overall integrity of the p53 pathway.

• Functional Assay of p53: This yeast-based assay provides a powerful and rapid assessment of p53's ability to transactivate its target genes. When human p53 is expressed in yeast, functional p53 can activate reporter genee.g., Ade2), leading to the synthesis of adenine and the formation of white colonies. Conversely, non-functional p53 mutants fail to activate Ade2, resulting in red colonies due to adenine auxotrophy. This qualitative assay offers a clear readout of p53's transcriptional competence.
• ELISA-based Assays: Utilizing enzyme-linked immunosorbent assay (ELISA) technology, these assays quantify the binding of active p53 to specific DNA response elements in nuclear extracts. This directly measures the DNA-binding capacity of p53, a prerequisite for its transcriptional regulatory functions.
• Luciferase Reporter Assays: These highly sensitive assays employ reporter gene constructs, typically luciferase, linked to p53 response elements. Upon p53 binding and activation, the reporter gene is transcribed and translated, producing a quantifiable luminescence signal. The intensity of this signal directly correlates with the level of p53 transcriptional activity, providing a quantitative measure of its functional integrity.

Immunohistochemistry (IHC) serves as a valuable complementary tool for assessing p53 protein expression in tissue samples. While wild-type p53 typically exhibits a short half-life and low cellular abundance, many mutant p53 proteins are conformationally altered, leading to increased stability and subsequent accumulation within the cell. This overexpression of mutant p53, detectable by IHC, often correlates with the presence of TP53 mutations and can provide a rapid, visual indication of p53 status in tumor biopsies.

For comprehensive and scalable analyses, Creative Biolabs integrates advanced high-throughput and multiplexed assay platforms:

• Bead-based multiplex Assay: This powerful platform enables the simultaneous detection and quantification of multiple proteins within the intricate p53 signaling network. By analyzing p53 alongside key interacting partners and downstream effectors, the assay provides a more comprehensive, pathway-level understanding of p53 activity and its regulatory context.

Creative Biolabs' extensive cell panel screening capabilities are central to our p53 service, offering a robust system for evaluating drug responses in the context of defined p53 statuses.

• Extensive Cell Line Library: We maintain a meticulously curated library of over 600 cancer cell lines, including engineered lines, representing a diverse array of cancer types and genetic backgrounds, including those with known TP53 mutations or wild-type p53. This vast resource allows for highly relevant in vitro drug testing.
• 2D and 3D Viability Assays: Our screening incorporates both conventional 2D monolayer cultures and more physiologically relevant 3D spheroid models, enabling a more accurate assessment of drug efficacy in a tumor microenvironment-mimicking context.
• Specialized Panels: For in-depth mechanistic studies, we offer specialized cell panels targeting critical pathways frequently dysregulated in cancer, such as DNA Damage Response (DDR), RAS, EGFR, and HER2. These panels facilitate a deeper understanding of drug mechanisms of action and drug sensitivity in specific genetic contexts.