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ab31900 is a recombinant protein produced in E. coli.
What are Affibody Molecules?
Affibody® affinity ligands 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. The current Affibody® libraries contains billions of variants. In function, Affibody® molecules mimic antibodies, nature’s own binders to an infinite number of antigens. Compared to antibodies, the most striking dissimilarity of Affibody® molecules is the small size. Affibody® molecules have a molecular weight of 14 kDa, compared to the molecular weight of antibodies, which is 150 kDa. In spite of its small size, the binding site of Affibody® molecules is similar to that of an antibody. The advantages of Affibody® molecules over antibodies are · their small size · the simple structure of the molecules · its robust physical properties · its ability to fold correctly intracellularly · the fast and cost-efficient production in bacteria · the possibility to produce Affibody® molecules through chemical synthesis · the possibility to couple Affibody® molecules in multimeric constructs.
This Anti-IgG Affibody® Molecule is modified with a unique C-terminal cysteine for directed single-point chemical modification, facilitating labelling with fluorescent dyes, biotin or 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. 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 ab31900 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.|
|Other||Use at an assay dependent concentration.
This product is also suitable for depletion of IgG from human serum and can be used as a detection reagent in a variety of assays.
SDS-PAGE analysis of flow-through fractions (FT) and eluted fractions after repeated affinity removal of IgG from human serum.
Lane 1: Untreated 5x diluted serum sample;
Lane 2: FT run 1;
Lane 3: FT run 75;
Lane 4: FT run 150;
Lane 5: FT run 225;
Lane 6: FT run 300;
Lane 7: eluate run 1;
Lane 8: eluate run 300;
Lane 9: IgG standard.