Anti β Amyloid  

Immunohistochemical detection of beta Amyloid using Anti-Beta Amyloid Antibody on TG APP23 mouse brain cortex frozen sections. Anti-Beta Amyloid Antibody used at 1:200 and incubated for 2 hours in TBS/BSA with Tween and azide. Fluorescent labelled anti rabbit IgG was then added. Carl Hobbs, King`s College London, United Kingdom.

Immunofluorescence microscopy of Rabbit Anti-Beta Amyloid antibody using HeLa cells fixed with MeOH. Anti-Beta Amyloid Antibody was used at 1 μg/mL, O/N at 4°C. Secondary antibody: Anti-RABBIT IgG DyLight？ 488 Conjugated Preadsorbed (p/n 611-741-127) at 2 ug/ml for 1 h at RT. Localization: APP is a cell surface protein that rapidly becomes internalized to endosomes and lysosomes. Some APP accumulates in secretory transport vesicles. Colocalizes with other proteins in a vesicular pattern in cytoplasm and perinuclear regions. Staining: Amyloid beta as green fluorescent signal with DAPI (blue) nuclear counterstain.

Immunofluorescence of Anti-Beta Amyloid Antibody. Tissue: adult mouse brain cells. Fixation: 4% paraformaldehyde. ？Antigen retrieval: none. Primary Antibody: Anti-Beta Amyloid diluted 1:20, 1:50, 1:100, and 1:200 in 0.1 M PBS-BSA-PLL ON at RT. Secondary Antibody: goat anti-rabbit AF488 conjugated antibody diluted 1:500 in 0.1 M PBS for 1 h at RT. Staining of beta-amyloid positive cells at 40x magnification. Independently Validated by？antibodies-online GmbH (p/n ABIN1043866/ ABIN95037) courtesy of Prof. Merighi, University of Turin.

Immunohistochemistry with anti-beta amyloid antibody showing amyloid beta plaque staining in human Alzheimer’s disease brain at 10x and 20x (B & C). Staining was performed on Leica Bond system using the standard protocol. Formalin fixed/paraffin embedded tissue sections were subjected to antigen retrieval with E1 (Leica Microsystems) retrieval solution for 20 min and then incubated with rabbit anti-beta amyloid antibody 600-401-253 at 1:100 dilution for 60 minutes. Biotinylated Anti-rabbit secondary antibody was used at 1:200 dilution to detect primary antibody. The reaction was developed using streptavidin-HRP conjugated compact polymer system and visualized with chromogen substrate, 3’3-diamino-benzidine substrate (DAB). The sections were then counterstained with hematoxylin to detect cell nuclei.

Human Heart (formalin-fixed, paraffin-embedded) stained with Anti-Beta Amyloid Antibody at 5 ug/ml followed by biotinylated goat anti-rabbit IgG secondary antibody, alkaline phosphatase-streptavidin and chromogen.

Western Blot of Rabbit anti-Beta Amyloid antibody. Marker: Opal Pre-stained ladder (p/n MB-210-0500). Lane 1: HEK293 lysate (p/n W09-000-365). Lane 2: HeLa Lysate (p/n W09-000-364). Lane 3: MCF-7 Lysate (p/n W09-000-360). Lane 4: Jurkat Lysate (p/n W09-000-370). Lane 5: A431 Lysate (p/n W09-000-361). Lane 6: LNCaP Lysate (p/n W09-001-GJ9). Lane 7: A-172 Lysate (p/n W09-001-GL5). Lane 8: NIH/3T3 Lysate (p/n W10-000-358). Load: 35 μg per lane. Primary antibody: Beta Amyloid antibody at 1:5,000 for overnight at 4°C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:30,000 for 60 min at RT. Blocking Buffer: 1% Casein-TTBS for 30 min at RT. Predicted MW: ~40-50 kDa. Observed size: ~48 kDa for Beta Amyloid.

Western Blot of Rabbit Anti-Beta Amyloid Antibody. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: HEK293T Whole Cell Lysate (p/n W09-001-GX5). Lane 3: Mouse Brain Whole Cell Lysate (p/n W10-000-004). Lane 4: A-172 Whole Cell Lysate (p/n W09-001-GL5). Lane 5: Daudi Whole Cell Lysate (p/n W09-001-MQ2). Load: 10μg/lane. Primary Antibody: Anti-Beta Amyloid at 1:1000 overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG HRP Conjugated (p/n 611-103-122) at 1:70,000 for 30min at RT. Block: BlockOut Buffer (p/n MB-073). Predicted MW: ~40-50kDa. Observed MW: ~48kDa.

Western Blot of Rabbit Anti-Beta Amyloid Antibody. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: HEK293T Whole Cell Lysate (p/n W09-001-GX5). Lane 3: Mouse Brain Whole Cell Lysate (p/n W10-000-004). Lane 4: A-172 Whole Cell Lysate (p/n W09-001-GL5). Load: 10μg/lane. Primary Antibody: Anti-Beta Amyloid at 1μg/mL overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG HRP Conjugated (p/n 611-103-122) at 1:70,000 for 30min at RT. Block: BlockOut Buffer (p/n MB-073). Predicted MW: ~40-50kDa. Observed MW: ~48kDa.

ELISA results of purified Rabbit anti-Beta Amyloid Antibody tested against BSA-conjugated peptide of immunizing peptide. Each well was coated in duplicate with 0.1μg of conjugate. The starting dilution of antibody was 5μg/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% fish gel, Goat anti-Rabbit IgG Antibody Peroxidase Conjugated (Min X Bv Ch Gt GP Ham Hs Hu Ms Rt & Sh Serum Proteins) (p/n 611-103-122) and TMB ELISA Peroxidase Substrate (p/n TMBE-1000).

Unfolding profile of Anti-Beta Amyloid Antibody. The fluorescence signal is plotted against temperature. The vertical line indicates the Ti at 83.8 °C. Independently Validated by？antibodies-online GmbH (p/n ABIN1043866/ ABIN95037) courtesy of NanoTemper Technologies.