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Immunohistochemistry with anti beta amyloid antibody showing amyloid beta plaque staining in human Alzheimers 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, 33 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.
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 ug/mL, O/N at 4C. Secondary antibody: Anti-RABBIT IgG DyLightTM 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.
ELISA results of purified Rabbit anti-Beta Amyloid Antibody tested against BSA-conjugated peptide of immunizing peptide. Each well was coated in duplicate with 0.1ug 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).
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: 10ug/lane. Primary Antibody: Anti-Beta Amyloid at 1ug/mL overnight at 2-8C. 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). Lane 5: Daudi Whole Cell Lysate (p/n W09-001-MQ2). Load: 10ug/lane. Primary Antibody: Anti-Beta Amyloid at 1:1000 overnight at 2-8C. 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. 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 ug per lane. Primary antibody: Beta Amyloid antibody at 1:5,000 for overnight at 4C. 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.
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