Amyloid beta oligomer-specific monoclonal antibody (OMAB), Biotinylated
AS10 932B | Clonality: Monoclonal | Host: Mouse | Reactivity: amyloid beta
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coating antibody at 2 µg/ml (ELISA), 1: 500 (IHC)
|Expected | apparent MW||4.5 kDa|
|Confirmed reactivity||human Abeta oligomers only
|Not reactive in||
no confirmed exceptions from predicted reactivity known in the moment
OMAB antibody is a versatile tool within research of Alzheimer’s disease. A sandwhich ELISA illustrates its potential regarding its high selectivity towards Aβ oligomers.
|Selected references||Richman et al. (2013). In Vitro and Mechanistic Studies of an Anti-Amyloidogenic Self-Assembled Cyclic D,L-#-Peptide Architecture. J. Americal Chemical Societ, Jan 19.
Lindhagen-Persson et al. (2010). Amyloid-β Oligomer Specificity Mediated by the IgM Isotype – Implications for a Specific Protective Mechanism Exerted by Endogenous Auto-Antibodies. PLoS ONE.
Abeta oligomer-specific antibody was adsorbed to Nunc-Immuno MaxiSorp plates (Nunc, Roskilde, Denmark) at 2 ug/ml in PBS. 1 ml of a 10 uM Aβ(1-42) sample containing a small fraction of Aβ-oligomers was separated using a superdex G75 (10/30) column. Aβ-fractions collected from the SEC were allowed to bind to OMAB plates for 20 minutes at 0°C. All fractions were analyzed and bound Aβwas detected using a polyclonal rabbit anti-Aβ antibody (AS08 328), Agrisera AB, Vännäs, Sweden) at a 1:1000 dilution followed by an anti-rabbit HRP-conjugated secondary antibody at a 1:5000 dilution (GE healthcare). ECBlue (Medicago, Uppsala, Sweden) was used as a substrate for HRP and the signal was detected by measuring the absorbance at 450 nm. Blocking solution and antibody-dilutions were made with 5% Non-fat dry milk in PBST and all washes were performed with PBS containing 0.1% Tween-20 (PBST).
10 µm of coronal sections from fresh-frozen transgenic mouse brain mutant (A) and wild type (B). Post-fixation in 4% formaldehyde solution, 5 min. OMAB antibody diluted 1:500, incubation at 4ºC ON. Mouse on mouse HRP-Polymer kit according to company instructions. Biocare Medical: BC-MM510 (Histolab) DAB substrate kit for peroxidase. Vector Laboratories: SK-4100 (ImmunKemi) Counterstained with Mayers HTX.
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