Alzheimer’s disease (AD) is a progressive, irreversible neurological disorder that destroys memory, thinking, and eventually, the ability to carry out simple tasks. The symptoms are mild at first, but worsen over time, and an affected person’s behavior becomes increasingly impulsive or unpredictable. One of the most characteristic features of AD is the formation of abnormal protein clumps, called amyloid plaques, and the presence of tangled bundles of fibers, known as neurofibrillary or tau tangles. These plaques and tangles collect between neurons and disrupt their normal function, thereby bringing about AD pathology. Neuronal damage in AD first appears to take place in the hippocampus, the region of the brain responsible for forming memories. With time, neuronal death occurs in other parts of the brain, ultimately leading to significant shrinkage of brain tissue.

While Alzheimer’s disease is one of the most common forms of dementia, there remains a lack of understanding of the pathogenesis of the disease. Research tools, such as antibodies, can help identify faulty proteins and decode why they behave differently and pave the way for potential therapeutics. Given the nature of the proteins known to be involved in AD, it becomes imperative that the antibodies scientists use to study them are highly target-specific and are robust enough to fish out proteins that are expressed in low quantities.

Alzheimer’s disease antibody targets:

Applications

14-3-3 beta/epsilon/zeta Antibody in Western Blot. Western blot was performed using 14-3-3 beta/epsilon/zeta Monoclonal Antibody (3C8) (Cat. No. MA1-34561, 1 µg/mL) and a 28 kDa band corresponding to 14-3-3 protein beta/epsilon/zeta was observed across the cell lines and tissues tested. Membrane enriched extracts (30 µg lysate) of MCF7 (Lane 1), HeLa (Lane 2), PC-3 (Lane 3), A549 (Lane 4), mouse skeletal muscle (Lane 5), rat skeletal muscle (Lane 6), mouse brain (Lane 7), and rat brain (Lane 8) were electrophoresed using NuPAGE 4-12% Bis-Tris Protein Gel (Cat. No. NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane by the iBlot 2 Gel Transfer Device (Cat. No. IB21001). The blot was probed with the primary antibody mentioned above and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal Recombinant Secondary Antibody, HRP (Cat. No. A28177, 1:10,000 dilution) using the iBright Imaging System (Cat. No. A44115). Chemiluminescent detection was performed using Novex ECL Chemiluminescent Substrate Reagent Kit (Cat. No. WP20005).

Tau Antibody (Cat. No. AHB0042) in IHC (F). Immunofluorescent analysis of p-Tau (S396) and Tau in human iPSC-derived forebrain organoids derived at day 40. The organoids were fixed with 4% PFA for 1 hour at room temperature, followed by incubation with 30% sucrose solution overnight at 4°C. The organoids were then embedded in OCT and cryosectioned at 5 µm, permeabilized with 0.2% Triton X-100 for 20 minutes and blocked with 10% donkey serum in PBS for 30 minutes at room temperature. Organoid slices were stained with Tau Monoclonal Antibody (TAU-5) (green; Cat. No. AHB0042) at a dilution of 1:500 and Phospho-Tau (Ser396) Polyclonal Antibody (red; Cat. No. 44-752G) at a dilution of 1:500 in blocking buffer overnight at 4°C. Then they were incubated with Donkey anti-Mouse IgG (H+L) ReadyProbes Secondary Antibody, Alexa Fluor 488 (Cat. No. R37114), Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 568 (Cat. No. A10042) at a dilution of 1:1000, as well as, DAPI (blue; 1:25000) in blocking solution at room temperature for 1 hour. Images were taken at 20X magnification. Scale bar: 50 µm.

Resources

For Research Use Only. Not for use in diagnostic procedures.