Clusterin Canine, HEK
Description
Production and Purification in HEK293 Cells
Clusterin Canine, HEK is produced via transient transfection in HEK293 cells, followed by chromatographic purification . The process involves:
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Transfection: HEK293 cells are transfected with a plasmid encoding the canine clusterin gene fused to a Flag tag .
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Secretion and Harvest: The protein is secreted into culture medium and purified using affinity chromatography (e.g., anti-Flag antibodies) .
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Lyophilization: The purified protein is filtered (0.4 µm) and lyophilized in Tris buffer (pH 7.5) .
This method ensures high yield and consistency, critical for research and diagnostic applications .
Biological Functions and Research Applications
Clusterin Canine, HEK exhibits diverse roles in canine physiology and pathology:
Apoptosis Regulation
Clusterin modulates apoptosis by binding to pro-apoptotic proteins, protecting cells from programmed death . In canine lymphoma, serum clusterin levels are significantly lower in treatment-naïve multicentric lymphoma (MLSA) dogs compared to healthy controls (p < 0.001) .
Tubular Injury Biomarker
In AKI, urinary clusterin (uClust) levels are elevated in dogs with IRIS AKI grades II–V, correlating with disease severity . For example:
Table 2: Urinary Clusterin in Canine AKI
Immunoregulatory Roles
Clusterin interacts with immune cells, influencing complement activation and phagocytosis . Its dysregulation is implicated in lymphoma and autoimmune diseases .
Lymphoma Biomarker
While serum clusterin levels in treatment-naïve MLSA dogs are lower than in healthy dogs, no significant difference is observed during remission . This variability limits its utility as a standalone diagnostic marker but suggests potential for monitoring treatment response .
AKI Prognosis
Urinary cystatin B (uCysB) and uClust are superior to traditional markers like SDMA for AKI prognosis . Dogs with non-surviving AKI show higher initial uCysB levels (731 ng/mL vs. 25 ng/mL in survivors, p = 0.009) .
Limitations
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Variability: Wide inter-individual differences in serum clusterin levels complicate interpretation .
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Cross-Species Reactivity: HEK-produced clusterin may differ in post-translational modifications compared to native canine proteins .
Research Priorities
Product Specs
Product Science Overview
Clusterin Canine Recombinant is produced in HEK293 cells (Human Embryonic Kidney 293 cells), which are widely used in biotechnology for the production of recombinant proteins. The recombinant protein is a single, glycosylated polypeptide chain containing 436 amino acids and has a molecular mass of 50.72 kDa . The protein is fused with a 13 amino acid Flag tag at the N-terminus, which aids in its purification and detection .
Clusterin is involved in several critical biological functions:
- Lipid Transport: It assists in the transport of lipids in the bloodstream.
- Cell Apoptosis: It plays a role in the regulation of programmed cell death.
- Tissue Remodeling: It is involved in the remodeling of tissues during development and in response to injury.
In research and biotechnology, recombinant Clusterin is used to study these functions and to develop therapeutic strategies for diseases where Clusterin is implicated, such as neurodegenerative diseases, cancer, and cardiovascular diseases.
