Product Introduction
1.1 BioMag series
The "Bio" in the BioMag series is derived from the prefix of our company name "Biodragon", and "Mag" stands for "Magnetic". All products containing BioMag are polymer magnetic microsphere products. At present, the BioMag series is mainly based on antibody affinity method labeled protein IP kits. It includes six types of tag IP kits: Anti-His-tag, Anti-Myc-tag, Anti-HA-tag, Anti-Flag-tag, Anti-GFP-tag and Anti-RFP-tag. In the future, we will develop more BioMag polymer magnetic bead-related products based on customer demands.
1.2 His Tag
The His-tag peptide is generally 6 or 8 histidine segments. It can be attached to the N-terminal or C-terminal of the fusion protein. Due to its small molecular weight and ease of separation and purification, the HIS-tag fusion tag has many obvious advantages compared with other tags and is currently the most widely used fusion tag for purification. His-tagged polypeptides can specifically bind to nickel ions, and recombinant proteins with histidine tags can be adsorbed by nickel columns, thereby achieving the purpose of purification.
1.3 Anti-His antibody
Anti-His antibodies can be used to detect the expression and intracellular localization of proteins fused with His tags, and can also be used for the purification, qualitative or quantitative detection of His fusion expression proteins. The Anti-His of our company has a very high affinity and is suitable for conducting protein interaction experiments such as WB and IP.
1.4 Components of the Kit
Antibodies have specific binding to the corresponding tags. By conjugating antibodies onto Beads, the fusion proteins with the corresponding tags can be bound to the Beads. After steps such as removing impurity proteins with buffer solution, the fusion protein can be eluted from the Beads using denaturants or high-temperature treatment, etc. For ease of use, this kit provides a variety of reagents to facilitate customers' better conduct protein interaction experiments.
1.5 Description of Elution buffer
Elution Buffer D contains a high concentration of urea and detergent, which can fully elute the tag protein and ensure that only a small amount of antibodies fall off the magnetic beads. However, there may still be a small amount of target protein remaining on the magnetic beads. It can also be directly added to the SDS Loading Buffer in the kit and treated at a high temperature of 95-100℃ for 5-10 minutes. The target protein can be completely detached from the magnetic beads, but the antibody will also completely fall off. When conducting subsequent WB experiments, the species of the secondary antibody should be carefully selected.
1.6 Positive control protein
The kit provides a red fluorescent protein with His tag. This protein emits visible red light under natural light and can be used to verify the binding capacity of Beads and eliminate possible deviations in the operation process. This protein is eukaryotic expressed and is shown as three bands with different molecular weights on SDS-PAGE. The binding time, binding conditions and elution conditions of different proteins with His tags to Beads are similar and can be referred to in parallel. However, considering that the binding of proteins to Beads is concentration-dependent, the operating conditions for positive control proteins should not be blindly followed without change.
1.7 Properties of Magnetic Beads
Polymer magnetic beads are made by using high-molecular polymerization technology to perfectly combine superparamagnetic materials and high-molecular materials. They have a faster magnetic response while maintaining the good dispersion of microspheres, extremely low non-specific adsorption and more abundant binding sites. BioMag Anti-His Beads are prepared based on the principle of high affinity between Biotin and streptavidin (SA) proteins. Firstly, SA proteins are high-density coupled to BioMag magnetic beads, and then biotinylated His antibodies are non-covalently coupled to SA polymer magnetic beads. Thus, it avoids the inactivation of antibodies caused by chemical covalent coupling. Therefore, BioMag Anti-His Beads have a higher antibody coupling density and antibody specific activity. It can be used for protein interaction experiments such as IP and Co-IP of HIS-tagged proteins secreted and expressed in prokaryotic and eukaryotic cell lysates or culture supernatants. Magnetic beads can be separated through magnetic racks or automatic separation instruments. The method is simple, efficient and convenient.