Amyloid beta peptide (A-beta 40/42) Antibody (Biotin)

SKU: orb3167451

Description

Our Anti-Amyloid beta peptide (A-beta 40/42) mouse monoclonal primary antibody detects human and rat Amyloid beta peptide (A-beta 40/42), and is IgG. It is validated for use in ELISA, ICC, IHC-Frozen, IHC-Paraffin-embedded, IP, WB.

Images & Validation

• Tested Applications: ELISA, ICC, IHC, IP, WB
• Dilution Range: WB: 1:2000-1:5000; IHC: 1:500-1:2000; ICC: 1:100-1:500; IP: 1:200-1:1000; ELISA: 1:50-1:1000
• Reactivity: Human, Rat
• Application Notes:
  The biotinylated MOAB-2 antibody has been tested by IHC and is also expected to work in applications validated for the unlabelled antibody (M-1586-100) at same or higher dilutions: Western Blotting (WB), Immunohistochemistry (IHC), Immunohistochemistry/paraffin embedded IHC(P), Immunoprecipitation (IP), Immunofluorescence (IF), ELISA. Western Blotting: MOAB-2 has been tested in WB using purified synthetic beta-amyloid preparations and from transgenic mouse brain formic acid extracts (see Figure 1). Formic acid extraction/concentration is required for Western blot detection from extracts. Standard ECL detection systems. Tissue samples for the detection of beta-amyloid should be prepared as detailed in Youmans KL et al. 2011 (Journal of Neuroscience Methods 196: 51-59) for best results. Detection of beta-amyloid 40/42 in direct westerns can be difficult; Dot-blots of prepared samples are recommended as detailed in Youmans KL et al. 2012. Immunohistochemistry: IHC:` Fresh frozen, 4% paraformaldehyde fixed frozen, or formalin fixed paraffin embedded tissues are all suitable. Antigen retrieval is required in fixed tissues for optimal staining. Antibody was tested on 4% paraformaldehyde/0.1% glutaraldehyde fixed frozen tissue from 3xTg and 5xFAD mice. MOAB-2 antibody detects intraneuronal and extracellular beta-amyloid in IHC and does not detect APP (Youmans KL et al. 2012). The antibody also reacts with archival formalin-fixed, paraffin-embedded tissue samples with antigen Heat Induced Epitope Retrieval (HIER). Recommended buffer for HIER is citrate, pH 6.0. Signal was weak without antigen retrieval. Immunoreactivity was observed in intraneural-amyloid deposition (plaque) in Alzheimer's brain. MOAB-2 was found to be extremely clean and with an excellent signal to noise ratio with no neuro-cellular diffusive staining. In addition, MOAB-2 demonstrated no significant differences in A-beta detection using paraffin fixed, free-floating sections (Youmans KL et al. 2012). Formic acid (FA) treatment resulted in optimal detection of both intraneuronal and extracellular A-beta compared to without FA (incubated in 88% FA 8 min, Youmans KL et al. 2012). Free floating tissue sections were permeabilized in TBS containing 0.25% Triton X-100 (TBSX; 3 x 10 min), blocked with 3% horse serum in TBSX (3 x 10 min) followed by 1% horse serum in TBSX (2 x10 min) and incubated with appropriate primary antibodies diluted in TBSX containing 1% horse serum overnight. See Youmans KL et al. 2012, for full IHC(P) protocol and method details. Immunofluorescence: IF: The antibody was tested on 4% PFA fixed frozen tissue. Fixed tissues were washed in TBS (3 x 10 min), then incubated in 88% FA (8 min), and then permeabilized in TBSX (3 x 10 min), and blocked in TBSX containing 5% rAlbumin (BSA; 1 hr). Sections were subsequently incubated with appropriate primary antibodies diluted in TBSX containing 2% BSA overnight on an oscillatory rotator. Detection was via fluorescently labelled absorbed secondary antibodies (Youmans KL et al. 2012). Immunoprecipitation: Labelled beta-amyloid using SA-coated beads as the capture vehicle, similar to the protocols employed by Youmans KL et al. 2012. ELISA: The antibody has been tested in ELISAs on synthetic beta-amyloid and tissue homogenates from beta-amyloid-Tg mice.

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Key Properties

• Host: Mouse
• Clonality: Monoclonal
• Clone No.: MOAB-2
• Immunogen: Recombinant human amyloid beta protein 42 (Aβ42): DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA
• Target: Amyloid beta peptide (A-beta 40/42)
• Conjugation: Unconjugated

Storage & Handling

• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Form/Appearance: Lyophilized
• Buffer/Preservatives: Lyophilized from PBS buffer, pH 7.4; contains no preservative.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

Beta-APP42; Beta-APP40; Beta-amyloid protein 42; Beta-amyloid protein 40; ABPP; APPI; Amyloid beta A4 protein; MOAB2; MOAB-2; Alzheimer's antibody; AB40; AB42; abeta

Quick Database Links

Gene Symbol

Amyloid beta peptide (A-beta 40/42)

UniProt

P05067