Anti-IgG Affibody®分子(ab31900)

概述

  • 产品名称Anti-IgG Affibody®分子
    参阅全部 IgG affibody®分子
  • 特异性ab31900 recognises the Fc part of IgG from several species with similar binding preferences as Protein A in terms of subclass specificities. This molecule binds with high affinity to human IgG of IgG1, IgG2 and IgG4 subclasses which comprise 92-98% of total IgG in a normal individual.
  • 经测试应用适用于: Othermore details
  • 种属反应性
    与反应: Mouse, Rabbit, Human, Rhesus monkey
  • 免疫原

    -

  • 常规说明

    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.

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
  • 存储溶液pH: 7.4
    Constituents: 0.079% Ammonium bicarbonate, PBS
  • Concentration information loading...
  • 纯化说明ab31900 is >98% pure, as determined by SDS-PAGE (Coomassie blue staining) and RP-HPLC analyses.
  • Affibody®分子说明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
  • 研究领域
  • 细胞定位Secreted
  • 别名
    • Ig gamma 1 chain C region
    • IgG
    • IGHG1
    • Immunoglobin heavy constant gamma 1
    • Immunoglobulin G
    see all

应用

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.

应用 Ab评论 说明
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.

Anti-IgG Affibody® Molecule 图像

  • Overlay chromatograms of repeated affinity removal of IgG from serum are shown. The chromatograms represent run number 1, 50 and 300 after consecutive injections of 700 ul of five times diluted human serum on 0.37 ml SulfoLink® Coupling Gel with immobilized Anti-IgG Affibody® molecule. The peak area of eluted fraction after each run is plotted in the image below. The identical chromatograms and consistent peak areas of eluted fractions prove that the depletion procedure can be reproducibly repeated at least 300 times without loss of binding capacity. SDS-PAGE analysis of flow-through fractions and eluted fractions shown in the third image demonstrate that the high specificity of the Anti-IgG Affibody® molecule is maintained through all the 300 consecutive injections. The capacity of this coupling gel allows for depletion of IgG from 1900 ul of five times diluted human serum per ml gel, corresponding to 380 ul of undiluted human serum per ml gel.
  • Peak area of the eluted fraction after each run of affinity removal. The consistent peak area prove that the depletion procedure can be reproducibly repeated at least 300 times without loss of binding capacity.
  • 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.

Anti-IgG Affibody® Molecule (ab31900)参考文献

This product has been referenced in:
  • Luckett BS  et al. Arcuate nucleus injection of an anti-insulin affibody prevents the sympathetic response to insulin. Am J Physiol Heart Circ Physiol 304:H1538-46 (2013). Read more (PubMed: 23542919) »

See 1 Publication for this product

Product Wall



The anti-IgG Affibody® molecule do contain 11 lysines in total. One is present in the binding site. If the intention is to perform coupling to the lysine, please be aware of potential decrease of binding performance.



Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"