COL4A3 Human, Biotin
Greater than 87% as determined by Capillary Electrophoresis.
Description
Human α3 chain of collagen IV; identical with the antigen called "glomerular basal membrane antigen" (GBM). Calculated Molecular weight: 48kDa. Calculated isoelectric point: pH 8.9. cDNA coding for a minicollagen version of the human collagen IV α3 chain fused to a 6-histidine purification tag. The term minicollagen designates the removal of most of the epitope-less triplehelical collagenous region (situated between the N-terminal 7S domain and the C-terminal noncollagenous NC1 domain), which is a requirement for recombinant production of this antigen.
Product Specs
This product represents the human alpha3 chain of collagen IV, also known as the glomerular basement membrane antigen (GBM). Its estimated molecular weight is 48kDa, and its calculated isoelectric point is pH 8.9. This product is a cDNA sequence that codes for a shortened version of the human collagen IV alpha3 chain. This 'minicollagen' version has most of its triple-helical collagenous region removed (this region lies between the N-terminal 7S domain and the C-terminal noncollagenous NC1 domain) to facilitate recombinant production. It also includes a 6-histidine purification tag fused to the minicollagen.
This product, COL4A3 Human biotin, is supplied in a solution containing 20mM HEPES (pH 7.5), 0.02% SDS, and 0.01mM EDTA.
For optimal short-term storage (2-4 weeks), keep the product refrigerated at 4°C.
For long-term storage, keep the product frozen at -20°C.
To maintain product integrity, avoid repeated freezing and thawing.
Analysis by Capillary Electrophoresis indicates a purity greater than 87%.
1. This product can bind to human autoantibodies of the IgG type.
2. It can be used in functional Streptavidin-based ELISA tests for the analysis of positive and negative samples.
Product Science Overview
Collagen Type IV Alpha 3, also known as COL4A3, is a crucial component of the basement membrane, a specialized structure that supports and separates different tissue types in the body. This protein is part of the larger collagen family, which is essential for maintaining the structural integrity of various tissues. The human recombinant, biotinylated version of Collagen Type IV Alpha 3 is a modified form of the protein that has been engineered for specific research and diagnostic applications.
Type IV collagen is unique among collagens because it forms a network rather than fibrils. It is composed of three alpha subunits, which can be any combination of six different genes (alpha 1 through alpha 6). The alpha 3 subunit, encoded by the COL4A3 gene, plays a significant role in the formation of this network. The biotinylation of the recombinant protein allows for easy detection and purification in various experimental setups.
The preparation of human recombinant Collagen Type IV Alpha 3 involves several steps:
- Gene Cloning: The COL4A3 gene is cloned into an appropriate expression vector.
- Expression: The vector is introduced into a host cell system, commonly Sf9 insect cells, which are used to produce the protein.
- Purification: The protein is purified using techniques such as affinity chromatography, often facilitated by the biotin tag.
- Biotinylation: The protein is biotinylated either during or after purification to enable its use in various assays.
For industrial-scale production, the process is scaled up using bioreactors. The steps include:
- Large-Scale Fermentation: The host cells are cultured in large bioreactors under controlled conditions to maximize protein yield.
- Harvesting: The cells are harvested, and the protein is extracted.
- Purification: Advanced purification techniques, including multiple chromatography steps, are employed to achieve high purity levels.
- Quality Control: The final product undergoes rigorous quality control to ensure consistency and functionality.
Collagen Type IV Alpha 3 undergoes several chemical reactions that are crucial for its function:
- Triple Helix Formation: The alpha subunits form a triple helix, which is stabilized by hydrogen bonds and other interactions.
- Network Formation: The triple helices interact to form a complex network, providing structural support to the basement membrane.
- Biotin-Streptavidin Interaction: The biotinylated protein can bind to streptavidin with high affinity, which is useful in various biochemical assays.
The biotinylated human recombinant Collagen Type IV Alpha 3 is used in:
- Research: Studying the structure and function of basement membranes.
- Diagnostics: Developing assays for detecting autoantibodies in diseases such as Goodpasture syndrome.
- Therapeutics: Potential applications in tissue engineering and regenerative medicine.
