Recombinant
RabMAb

Recombinant Anti-Amyloid Fibril antibody [mOC22] - Conformation-Specific (ab205339)

Overview

  • Product name

    Anti-Amyloid Fibril antibody [mOC22] - Conformation-Specific
    See all Amyloid Fibril primary antibodies
  • Description

    Rabbit monoclonal [mOC22] to Amyloid Fibril - Conformation-Specific
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, Dot blotmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Other Immunogen Type corresponding to Human Amyloid Fibril. (Amyloid beta 1-42 fibrils).
    Database link: P05067

  • Positive control

    • Dot Blot: beta Amyloid (Aß) 1-40 and beta Amyloid (Aß) 1-42. IHC-P: FFPE Hu Brain Alzheimer.
  • General notes

    This antibody was developed as part of a collaboration between Abcam and Professor Charles Glabe, UC Irvine.

    ab205339 (mOC22)  recognizes a conformation-dependent and aggregation-specific generic fibril epitope that is independent of the peptide sequence.  Although it maps to a linear segment of Aß (residues 3-7, EFRHD) it also reacts with alpha synuclein and islet amyloid polypeptide (IAPP) fibrils but not monomers (Hatami et al 2014). mOC22 preferentially stains the central core of cored plaques. mOC22 also stains misfolded or aggregated intraneuronal amyloid deposits (Hatami et. al 2014).  Immunoreactivity on western blots is enhanced by boiling the membrane.

    For further information on the immunogen, please refer to Hatami et al. 2014 and Kayed et al. 2007.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab205339 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use a concentration of 0.1 - 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Dot blot 1/7000.

Target

  • Cellular localization

    Membrane.
  • Database links

  • Alternative names

    • AAA antibody
    • ABETA antibody
    • ABPP antibody
    • AD1 antibody
    • Alzheimer disease amyloid protein antibody
    • Amyloid beta precursor protein antibody
    • APP antibody
    • APPI antibody
    • Beta amyloid peptide antibody
    • Cerebral vascular amyloid peptide antibody
    • CTFgamma antibody
    • CVAP antibody
    • Peptidase nexin II antibody
    • PN II antibody
    • PN2 antibody
    • PreA4 antibody
    • Protease nexin II antibody
    see all

Images

  • IHC image of Amyloid Fibril staining in Human Brain Alzheimer formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab205339, 0.1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Dot blot analysis of beta Amyloid labeled with ab205339 at 1/7000 dilution.

    Lane 1: beta Amyloid (Aβ) 1-40;

    Lane 2: beta Amyloid (Aβ) 1-42.

    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/30000 was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 30 seconds.

    Note: Antibody reactivity was assessed using a dot blot, which is a non-quantitative method that maintains the native conformation of beta Amyloid.  beta Amyloid 1-40 and 1-42 peptides underwent the following aggregation conditions before being spotted onto a nitrocellulose membrane and detected using ab205339:

    Monomers: 0.3 mg of beta Amyloid peptide was dissolved in 30 µl 100 mM NaOH and incubated at room temperature for 10 minutes. It was then diluted with 970 µl of 1% SDS and boiled for five minutes.

    Oligomers: 0.3 mg of beta Amyloid peptide was dissolved in 30 µl 100 mM NaOH and incubated at room temperature for 10 minutes. It was then diluted with 970 µl of 10 mM phosphate buffer pH 7.4 containing 0.02% sodium azide and incubated at room temperature for four days.

    Fibrils: 0.3 mg of beta Amyloid peptide was dissolved in 1 ml 50% hexafluoroisopropanol (HFIP) with 0.02% sodium azide. It was then stirred constantly for nine days; the first seven with a cap on and the final two with the cap removed to allow evaporation of the HFIP. Fibrils were then sedimented at 20,000 rpm in a microcentrifuge for 20 minutes and resuspended in 1 ml of PBS + 0.02% sodium azide.

  • Negative control (secondary ab only) Dot blot analysis of beta Amyloid.

    Lane 1: beta Amyloid (Aβ) 1-40;

    Lane 2: beta Amyloid (Aβ) 1-42.

    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/30000 was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 30 seconds.

References

This product has been referenced in:

  • Hatami A  et al. Familial Alzheimer's Disease Mutations within the Amyloid Precursor Protein Alter the Aggregation and Conformation of the Amyloid-ß Peptide. J Biol Chem 292:3172-3185 (2017). Read more (PubMed: 28049728) »
  • Hatami A  et al. Monoclonal antibodies against Aß42 fibrils distinguish multiple aggregation state polymorphisms in vitro and in Alzheimer disease brain. J Biol Chem 289:32131-43 (2014). Read more (PubMed: 25281743) »
See all 2 Publications for this product

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