His Tag Antibody
Description
Monoclonal antibodies are produced by immunizing mouse with synthesized polypeptide 6 x His. This antibody was purified via protein-A affinity chromatography.
Product Specs
Monoclonal antibodies are generated by immunizing mice with a synthesized 6xHis polypeptide. Purification of this antibody is achieved through protein-A affinity chromatography.
Provided as a 1mg/ml solution in PBS (pH 7.4) containing 0.02% Sodium Azide and 10% Glycerol.
Suitable applications include ELISA and Western Blot (W.B.). Researchers are advised to determine the optimal dilutions for their specific experiments.
PAT1D6AT.
Kappa IgG1.
Product Science Overview
A polyhistidine tag, commonly referred to as a His-tag, is an amino acid sequence consisting of at least six histidine residues. This tag is often used in protein purification processes due to its strong affinity for metal ions, particularly nickel. The His-tag can be added to either the N- or C-terminus of a protein, facilitating its isolation and purification through immobilized metal ion affinity chromatography (IMAC) .
The principle behind the polyhistidine tag lies in the ability of histidine residues to coordinate with metal ions. When a His-tagged protein is introduced to a carrier with immobilized metal ions, such as nickel, the histidine residues chelate the metal ions, binding the protein to the carrier. This allows for the selective isolation of the His-tagged protein, as other proteins either do not bind or bind weakly and can be washed away. The tagged protein can then be eluted from the carrier using an appropriate buffer .
Polyhistidine tags are widely used in various applications, including:
- Protein Purification: His-tags facilitate the purification of recombinant proteins by allowing for easy separation from other cellular components.
- Protein Detection: His-tagged proteins can be detected using anti-His antibodies in techniques such as Western blotting and ELISA.
- Protein Crystallization: The tag can aid in the crystallization of proteins for structural studies.
Mouse antibodies are immunoglobulins produced by mice in response to antigens. These antibodies are often used in research and therapeutic applications due to their specificity and ability to target particular antigens. However, the use of mouse antibodies in humans can lead to the development of human anti-mouse antibodies (HAMA), which can reduce the effectiveness of the treatment and cause adverse reactions .
When a polyhistidine tag is added to a mouse antibody, it allows for the efficient purification and detection of the antibody. This combination is particularly useful in research settings where the isolation of specific antibodies is required for further study or application. The His-tagged mouse antibodies can be purified using IMAC and detected using anti-His antibodies, streamlining the experimental process.
