Recombinant
RabMAb

Recombinant Anti-vascular Amyloid 1-42 antibody [mOC31] - Conformation-Specific (ab201059)

Overview

  • Product name

    Anti-vascular Amyloid 1-42 antibody [mOC31] - Conformation-Specific
    See all vascular Amyloid 1-42 primary antibodies
  • Description

    Rabbit monoclonal [mOC31] to vascular Amyloid 1-42 - Conformation-Specific
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Human
  • Immunogen

    Other Immunogen Type corresponding to Human vascular Amyloid 1-42. Amyloid beta 1-42 fibrils were used as the immunogen.
    Database link: P05067

  • Positive control

    • beta Amyloid (Aß) 1-42 IHC-P: FFPE human brain Alzheimer
  • General notes

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

    ab201059 recognizes a conformation-specific discontinuous epitope of beta Amyloid that maps to segments EFGRHSGY and ED (Hatami et al. 2014). It specifically recognizes a subset of vascular amyloid that does not colocalize with thioflavin S (Hatami et al. 2014, McLean et al. 2013).

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    mOC31
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab201059 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 1/100.
IHC-P 1/250 - 1/500. Do not perform antigen retrieval.
IHC-FoFr Use at an assay dependent concentration.
IHC-FrFl Use at an assay dependent concentration.

Target

  • Alternative names

    • A4 antibody
    • A4_HUMAN antibody
    • AAA antibody
    • ABETA antibody
    • ABPP antibody
    • AICD-50 antibody
    • AICD-57 antibody
    • AICD-59 antibody
    • AID(50) antibody
    • AID(57) antibody
    • AID(59) antibody
    • Alzheimer disease amyloid protein antibody
    • Amyloid intracellular domain 50 antibody
    • Amyloid intracellular domain 57 antibody
    • Amyloid intracellular domain 59 antibody
    • APP antibody
    • APPI antibody
    • Beta amyloid protein 42 antibody
    • Beta APP42 antibody
    • Beta-APP40 antibody
    • Beta-APP42 antibody
    • C31 antibody
    • Cerebral vascular amyloid peptide antibody
    • CVAP antibody
    • Gamma-CTF(50) antibody
    • Gamma-CTF(57) antibody
    • Gamma-CTF(59) antibody
    • PN-II antibody
    • PreA4 antibody
    • Protease nexin-II antibody
    • S-APP-alpha antibody
    • S-APP-beta antibody
    see all

Images

  • IHC image of beta Amyloid staining in a formalin fixed paraffin embedded human Alzheimer brain tissue section, performed on a Leica Bond™ system using the standard protocol F. No antigen retrieval was performed prior to staining. The section was incubated with ab201090 at 1/500 dilution 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.

     Image showing staining of vascular Amyloid as described in Hatami et al. 2014

  • Dot blot analysis of beta Amyloid 1-42 labeled with ab201059 at 1/100 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: 1 minute.

    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 ab201059:
    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 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 using ab201059 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.

  • PFA perfusion-fixed frozen sections of APP transgenic mouse brain stained for beta-Amyloid 1-42 with conformation-specific ab201059 (1/500) in immunohistochemical analysis (green). The nuclear counter stain is DAPI (blue).

    See Abreview

  • Negative control (secondary ab only):
    Lane 1: beta Amyloid (Aβ) 1-40.
    Lane 2: beta Amyloid (Aβ) 1-42.

    Primary antibody omitted.
    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:

  • 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 3 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
Yes - 0.5% Triton-X PBS
Specification
Brain
Fixative
Formaldehyde

Sumit Sarkar

Verified customer

Submitted Jun 20 2016

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - 0.1 M PBS with 1% Triton X
Specification
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 Feb 05 2016

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