This product is a recombinant protein produced in E.coli.
What are Affibody Molecules?
Affibody® affinity ligands are unique research reagents, produced using innovative protein-engineering technologies. They are small, simple proteins composed of a three-helix bundle based on the scaffold of one of the IgG-binding domains of Protein A. Protein A is a surface protein from the bacterium Staphylococcus aureus. This scaffold has excellent features as an affinity ligand and can be designed to bind with high affinity to any given target protein. The domain consists of 58 amino acids, 13 of which are randomized to generate Affibody® libraries with a large number of ligand variants. Thus, the libraries consist of a multitude of protein ligands with an identical backbone and variable surface-binding properties. In function, Affibody® Molecules mimic monoclonal antibodies. Compared to antibodies, the most striking dissimilarity of Affibody® Molecules is the small size. Affibody® Molecules have a molecular weight of 6kDa, compared to the molecular weight of antibodies, which is 150kDa. In spite of its small size, the binding site of Affibody® Molecules is similar to that of an antibody. The advantages of Affibody® Molcules over antibodies are: -their small size -the simple structure of the molecules -its robust physical properties; able to withstand a broad range of analytical conditions, including extreme pH and elevated temperature -its ability to fold correctly intracellularly -the fast and cost effective production in bacteria -the potential to couple Affibody® Molecules in multimeric constructs Affibody® Molecules have highly competitive properties for applications within affinity purification, sample preparation, protein detection and in vitro diagnostics.
This Anti-Fibrinogen Affibody® Molecule is modified with a unique C-terminal cysteine for directed single-point chemical modification, facilitating coupling to matrices. However, tail-to-tail dimers are spontaneously generated via a disulphide bridge between the C-terminal cysteines. Prior to coupling via the C-terminal the Affibody® Molecule needs to be reduced to expose the reactive cysteine residue. Recommended reducing condition is 20mM DTT at a pH above 7.5 and incubation at room temperature for 2 hours. Remove excess DTT by passage through a desalting column, not by dialysis. Not yet tested in other applications. Optimal dilutions/concentrations should be determined by the end user. ab50345 is a secondary antibody suitable for use in the process of detecting this Affibody® Molecule.
THIS AFFIBODY® MOLECULE REQUIRES CONJUGATION TO A SUITABLE LABEL BEFORE USE. PLEASE REFER TO THE "PROTOCOLS" LINK BELOW.
Our Abpromise guarantee covers the use of ab36058 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|AP||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|Other||Use at an assay dependent concentration.
This Anti-IgA Affibody® Molecule is an ideal affinity ligand as capture reagent in ELISA and as capture molecule in affinity chromatography.
Results quantitative ELISA
The Anti-IgA Affibody® molecule can be used as capture reagent in a sandwich ELISA in combination with a goat anti-IgA antibody as the detection reagent. Titration of IgA gives a sigmoid curve with a sensitivity of 0.2 ng IgA/ml (defined as two times background value).
Analysis of iga concentration in depleted serum
The remaining IgA in samples from serum, depleted from IgA by passage through an Anti-IgA Affibody® molecule coupled column, was analyzed using the Anti-IgA Affibody® ELISA. Concentration of IgA in flow through samples from 60 ul, 0.2 ml and 0.4 ml depleted plasma was analyzed and the percentage of achieved depletion was calculated. The data is presented in table 1.
ab36058 has not yet been referenced specifically in any publications.