Clusterin Mouse
CLI, AAG4, KUB1, SGP2, SGP-2, SP-40, TRPM2, MGC24903, Clusterin, Apolipoprotein J, Apo-J.
Filtered White lyophilized (freeze-dried) powder.
Greater than 90.0% as determined by SDS-PAGE.
Description
Clusterin Mouse Recombinant is a single, glycosylated polypeptide chain containing 433 amino acids (22-448a.a) and having a molecular mass of 50.2kDa (calculated). Clusterin is fused to a 6 a.a His tag at C-terminal.
Product Specs
Clusterin, also called Apolipoprotein J (APO-J), is a protein typically with a molecular weight of 75-80 kDa. It exists as two disulfide-linked chains, forming a heterodimer. This protein is heavily glycosylated, meaning it has sugar molecules attached, and these sugars make up around 30% of its structure. Interestingly, shorter versions of Clusterin have been found inside the cell nucleus. The process of creating Clusterin involves cutting a longer precursor protein. First, a 22-amino acid signal sequence, which guides the protein's production, is removed. Then, the precursor is split between amino acids 227 and 228, resulting in the two chains (a and b) that form the active Clusterin. These chains align in opposite directions and are held together by five disulfide bonds within regions rich in cysteine amino acids. The structure also includes coiled-coil and amphipathic alpha-helices, which are common protein structural motifs. Clusterin's amino acid sequence is highly conserved across different species, ranging from 70% to 80% similarity. It's found in almost all tissues of mammals and can be detected in various fluids like blood plasma, milk, urine, cerebrospinal fluid, and semen. Clusterin's ability to bind with other molecules is noteworthy. It interacts with immunoglobulins, lipids, heparin, bacteria, components of the complement system, paraoxonase, beta-amyloid, leptin, and others. This diverse binding repertoire suggests its involvement in a wide array of functions, including attracting immune cells, promoting aggregation, preventing complement attack, inhibiting cell death, remodeling membranes, transporting lipids and hormones, and scavenging harmful substances. The protein's activity is often altered in disease conditions. Clusterin levels, whether at the RNA or protein level, can increase or decrease in conditions like cancer, tissue repair, infections, Alzheimer's disease, retinitis pigmentosa, heart attack, kidney damage, autoimmune disorders, and more, highlighting its potential relevance in various disease mechanisms.
Recombinant Mouse Clusterin is a single-chain protein produced artificially, containing sugar modifications. It consists of 433 amino acids (specifically, positions 22 to 448 of the original protein sequence), resulting in a molecular weight of 50.2 kDa as calculated from its composition. A His tag, consisting of six histidine amino acids, has been added to the C-terminal end of the protein to aid in purification and detection.
The product appears as a white powder that has been sterilized by filtration and dried by freeze-drying.
The Clusterin protein has undergone filtration (using a 0.4 µm filter) and freeze-drying. It is supplied in a solution containing 20 mM Tris buffer and 50 mM NaCl at a pH of 7.5. The concentration before freeze-drying is 0.5 mg/ml.
To prepare a working solution, it is advised to add deionized water to the lyophilized powder to achieve a concentration of around 0.5 mg/ml. Ensure the powder completely dissolves to form a clear solution.
For long-term storage, keep the lyophilized protein at -20°C. After adding water to reconstitute the protein, divide it into smaller portions (aliquots) for future use. This minimizes the number of freeze-thaw cycles, which can damage the protein. Reconstituted protein can be stored at 4°C for a limited time. Stability studies show no significant changes in the protein after storage at 4°C for two weeks.
The purity of the Clusterin protein is greater than 90.0% as determined by SDS-PAGE, a common technique to separate and analyze proteins based on their size.
CLI, AAG4, KUB1, SGP2, SGP-2, SP-40, TRPM2, MGC24903, Clusterin, Apolipoprotein J, Apo-J.
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Product Science Overview
Apolipoprotein-J is involved in several critical biological processes, such as:
- Lipid Transport: ApoJ is a component of high-density lipoproteins (HDL) and is involved in lipid transport and metabolism.
- Tissue Remodeling: It participates in tissue remodeling and repair processes.
- Apoptosis: ApoJ has anti-apoptotic properties, protecting cells from programmed cell death.
In the context of neurodegenerative diseases, ApoJ is particularly noteworthy for its role in amyloid-β (Aβ) aggregation and clearance, which is relevant to conditions like Alzheimer’s disease.
Recombinant ApoJ can be produced using various expression systems, including bacterial, yeast, insect, and mammalian cells. The choice of expression system depends on the desired yield, post-translational modifications, and biological activity of the protein.
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Bacterial Expression System:
- Transformation: The gene encoding ApoJ is inserted into a plasmid vector and introduced into bacterial cells (e.g., E. coli).
- Induction: Protein expression is induced using agents like IPTG.
- Purification: The recombinant protein is purified using affinity chromatography techniques.
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Mammalian Expression System:
- Transfection: Mammalian cells (e.g., HEK293) are transfected with a plasmid containing the ApoJ gene.
- Expression: The cells are cultured, and the protein is expressed and secreted into the culture medium.
- Purification: The protein is purified from the culture medium using affinity chromatography.
ApoJ undergoes various post-translational modifications, including glycosylation, which can affect its function and stability. Analytical techniques such as mass spectrometry and Western blotting are used to characterize these modifications.
- Mass Spectrometry: This technique helps identify and quantify the post-translational modifications of ApoJ.
- Western Blotting: Used to detect and analyze the expression levels and modifications of ApoJ.
ApoJ’s role in amyloid-β aggregation and clearance is of particular interest in Alzheimer’s disease research. Studies have shown that recombinant ApoJ can reduce amyloid-β toxicity and aggregation in vitro, suggesting potential therapeutic applications .
In a transgenic mouse model of cerebral amyloid angiopathy, chronic treatment with recombinant human ApoJ (rhApoJ) has been shown to reduce the occurrence of cerebral microbleeds and improve neurovascular health . This highlights the potential of ApoJ as a therapeutic agent in neurodegenerative diseases.
