Anti-beta Amyloid 1-42 antibody [mOC64] - Low endotoxin, Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Amyloid 1-42 antibody [mOC64] - Low endotoxin, Azide free (AB224275)

This data was developed using ab201060, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human alzheimer brain labelling beta Amyloid with ab201060 at a concentration of 0.1μg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab201060 Anti-beta Amyloid 1-42 antibody [mOC64] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• Dot

Supplier Data

Dot Blot - Anti-beta Amyloid 1-42 antibody [mOC64] - Low endotoxin, Azide free (AB224275)

Dot blot analysis of beta Amyloid 1-42 labeled with ab201060 at 1/7000 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 ab201060 :
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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201060).

• Dot

Supplier Data

Dot Blot - Anti-beta Amyloid 1-42 antibody [mOC64] - Low endotoxin, Azide free (AB224275)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201060).