A 3 µL amount of 40 µM Aβ1–40 sample was spotted onto nitrocellulose membrane (Bio-Rad) at selected time points. The membranes were blocked by 10% (w/v) fat-free milk in 50 mM Tris 150 mM NaCl, pH 7.4, and 0.05% (v/v) Tween-20 (TBST buffer) for 1 h at room temperature, followed by incubation with antioligomer polyclonal A11 antibody (1:1000 dilution; Invitrogen) or Aβ1–16 (6E10) monoclonal antibody (1:1000 dilution; Covance) in 5% (w/v) fat-free milk and TBST buffer overnight at 4 °C. The membranes were washed 3 times in TBST before incubation with antirabbit or antimouse antibody (1:5000 dilution) in 5% (w/v) fat-free milk and TBST buffer at room temperature for 1 h.
A11 antibody
Manufactured by Thermo Fisher Scientific
The A11 antibody is a laboratory reagent used for research purposes. It is a monoclonal antibody that specifically recognizes and binds to a particular target protein or antigen. The core function of the A11 antibody is to serve as a detection or analytical tool in various research applications, such as immunoassays, Western blotting, or immunohistochemistry. The detailed specifications and intended use of this product are not available within the scope of this request.
Automatically generated - may contain errors
Lab products found in correlation
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Aβ1 16 6e10 monoclonal antibody, by Fortrea (1 mentions)
Superdex 200 10 300 column, by GE Healthcare (1 mentions)
Benzonase nuclease, by Merck Group (1 mentions)
0.45 μm nitrocellulose membrane, by Cytiva (1 mentions)
0.22 μm filter, by Merck Group (1 mentions)
Hrp conjugated anti rabbit secondary antibody, by Jackson ImmunoResearch (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
4 protocols using a11 antibody
1
Amyloid-beta Oligomer Detection
2
Characterization of Huntingtin Oligomers
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N2A cells were co-transfected with pcDNA3-Htt(Q)25-eYFP or pcDNA3-Htt(Q)109-eYFP and either pcDNA3 vector or pcDNA3.1-FKBP12–V5. After 48 hours, the transfected cells were harvested in 500 μl of ice-cooled PBS buffer containing protease inhibitor cocktail (Roche) and Benzonase Nuclease (Merck Millipore), and sonicated on ice for 1 min. Extracts were centrifuged at 14 K rpm for 30 min at 4 °C, and concentrations were determined using the BCA assay. Samples containing 120 μg of total proteins in a volume of 500 μl were filtered with a 0.22 μm filter (Millipore) and fractionated with a Superdex 200 10/300 column (GE Healthcare) using a flow rate of 0.3 ml/min. Each fraction (1 ml volume/fraction) was collected and subjected to Western blot and slot blot analysis. Htt oligomeric species in each fraction were quantified by densitometry (Image J). The percentage of Htt species was calculated by dividing the density of each fraction by the summed density of every fraction (fractions 7–18). For slot-blotting analysis, the collected fractions were applied to a 0.45 μm nitrocellulose membrane (Schleicher & Schuell) and probed with an A11 antibody (1:1000, Invitrogen) overnight at 4 °C, followed by incubation with HRP-conjugated anti-rabbit secondary antibody (1:7500; Jackson ImmunoResearch) for 1 hour at room temperature. Blots were detected using an ECL chemiluminescent kit.
3
Dot Blot for Prefibrillar Aβ Oligomers
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For detection of prefibrillar Aβ oligomers, 1 μg of protein was spotted on a nitrocellulose membrane and probed with A11 antibody (1 : 2000, Invitrogen). The membranes were probed with anti-rabbit secondary antibody, and immunoreactive signals visualized using enhanced chemiluminescence. The exact same number of samples was spotted on additional dot blots and probed with 6E10 antibody and Bradford reagent for normalization.
4
Curcumin Synthesis and Analogue Protocols
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All solvents used for the synthesis of curcumin and its analogues were either of
spectral grades or distilled prior to use. Solvents and other reagents for
synthesis were procured from Sigma Aldrich, USA unless otherwise mentioned.
Chemicals used for bacterial cell culture and mammalian cell cultures were
obtained from Himedia, India and Gibco, USA respectively. Antibodies against
α-synuclein were procured from Santa cruz Biotechnology Inc, USA
while A11 antibody was obtained from Invitrogen, CA, USA. All other chemicals
used were from Sigma Aldrich, USA.
spectral grades or distilled prior to use. Solvents and other reagents for
synthesis were procured from Sigma Aldrich, USA unless otherwise mentioned.
Chemicals used for bacterial cell culture and mammalian cell cultures were
obtained from Himedia, India and Gibco, USA respectively. Antibodies against
α-synuclein were procured from Santa cruz Biotechnology Inc, USA
while A11 antibody was obtained from Invitrogen, CA, USA. All other chemicals
used were from Sigma Aldrich, USA.
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