Recombinant
RabMAb

Recombinant Anti-Amyloid Fibril antibody [mOC87] - Conformation-Specific (ab201062)

Overview

  • Product name

    Anti-Amyloid Fibril antibody [mOC87] - Conformation-Specific
    See all Amyloid Fibril primary antibodies
  • Description

    Rabbit monoclonal [mOC87] to Amyloid Fibril - Conformation-Specific
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Dot blot, IHC-FoFr, IHC-P, IHC-FrFlmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human Amyloid Fibril. Amyloid beta 1-42 fibrils were used as the immunogen.
    Database link: P05067

  • Positive control

    • beta Amyloid (Aß) 1-40; beta Amyloid (Aß) 1-42. IHC-P: FFPE human Alzheimer hippocampus tissue sections.
  • General notes

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

    ab201062 recognizes a generic epitope of amyloid fibrils and oligomers that is independent of linear sequence (Hatami et al. 2014). Its reactivity with Aß monomer and oligomers is decreased or eliminated upon thermal denaturation at 100°C of Aß in SDS sample buffer on western blots (Hatami et al. 2014).

    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 ab201062 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
Dot blot 1/8000.
IHC-FoFr Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-FrFl Use at an assay dependent concentration.

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

  • Immunohistochemical staining of human brain tissue from a patient with a diagnosis of Alzheimers disease, male, 81 years, 5 hour post mortem index, tangle stage 5, plaque stage B, mini mental status exam score 12. Sections were cut using a vibratome. No antigen retrieval was performed. Free floating sections were stained using ab201062 at a dilution of 50 ng/mL. The secondary antibody used was a biotinylated goat anti-rabbit at a dilution of 1/225, which was blocked with normal goat serum. The sample was visualized using ABC solution (1 hour incubation) followed by 1-4 minutes of DAB. The sample was mounted and allowed to dry overnight, followed by dehydration in increasingly concentrated ethanol solutions.

  • Dot blot analysis of beta Amyloid 1-42 labeled with ab201062 at 1/8000 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 1/5000 dilution was used as secondary antibody.
    Blocking and diluting buffer: 5% NFDM/TBST.
    Exposure time: 30 seconds.

    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 ab201062:
    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.

     

  • Immunohistochemical analysis of 4% PFA in 0.1M PBS perfusion-fixed murine APP-PS1 transgenic brain tissue sections, labelling beta amyloid with ab201062 at a dilution of 1/200 incubated for 24 hours at 4°C in 0.1 M PBST with 10% donkey serum. Permeabilization was 0.1M PBS with 3% Triton X. Secondary was a polyclonal rabbit Alexa Fluor® 488 at 1/100. Counterstaining was DAPI against nuclear DNA and astrocytes stained with GFAP-Cy3.

    See Abreview

  • IHC image of beta Amyloid staining in human Alzheimer hippocampus 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 ab201062, 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

  • Negative control (secondary ab only):

    Lane 1: beta Amyloid (Aβ) 1-40.
    Lane 2: beta Amyloid (Aβ) 1-42.
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/5000 dilution was used as secondary antibody.
    Blocking and diluting buffer: 5% NFDM/TBST.
    Exposure time: 30 seconds.

References

This product has been referenced in:

  • Yu L  et al. The New Application of UHPLC-DAD-TOF/MS in Identification of Inhibitors on ß-Amyloid Fibrillation From Scutellaria baicalensis. Front Pharmacol 10:194 (2019). Read more (PubMed: 30936829) »
  • 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) »
See all 3 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - whole (Brain lysate)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
30 µg
Specification
Brain lysate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Nov 09 2015

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (APP-PS1 transgenic mouse brain)
Permeabilization
Yes - 0.1 M PBS with 3% Triton X
Specification
APP-PS1 transgenic mouse brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Nov 06 2015

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