Paraffin-embedded brain tissues were sectioned at 5-μm thickness. Deparaffinization was achieved by washing slides through a series of xylenes and decreasing concentrations of ethanol. Tissue sections were then subjected to antigen retrieval using citrate buffer, pH 6.0 (Sigma-Aldrich) at 95 °C for 30 min. Following rinsing with PBS, sections were incubated in blocking buffer containing PBS with 10% normal donkey serum and 0.03% Triton-X (Sigma-Aldrich) for 2 h at room temperature. Slides were then incubated with primary antibody in blocking buffer overnight at 4°C. The following day, slides were rinsed with PBS then incubated in appropriate Alexa Fluor secondary antibodies (Thermo Fisher Scientific). Sections were then rinsed and stained with Hoechst DNA dye (Thermo Fisher Scientific) before being coverslipped with ProLong mounting medium (Invitrogen). Primary antibodies included rat anti-human CD3 (Abcam), rabbit anti-human CD8α (Cell Signaling), mouse anti-Aβ (Cell Signaling), chicken anti-human MAP2 (Abcam), mouse anti-human granzyme-A (Abcam), rat anti-mouse CD8a (eBioscience) and rabbit anti-mouse NEFH (Abcam). For mouse Aβ plaque staining, ThioflavinS (1 mg ml−1, 1:625, Sigma) was added to the secondary antibody solution.
Mouse anti aβ
Manufactured by Cell Signaling Technology
Sourced in United States
Mouse anti-Aβ is a monoclonal antibody that specifically recognizes the amyloid-beta (Aβ) peptide. It is a laboratory tool used to detect and study Aβ in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Citrate buffer, by Merck Group (1 mentions)
Prolong mounting medium, by Thermo Fisher Scientific (1 mentions)
Triton x, by Merck Group (1 mentions)
Rat anti mouse cd8a, by Thermo Fisher Scientific (1 mentions)
Hoechst dna dye, by Thermo Fisher Scientific (1 mentions)
Alexa fluor secondary antibody, by Thermo Fisher Scientific (1 mentions)
Thioflavin s, by Merck Group (1 mentions)
L a b solution, by Polysciences (1 mentions)
Rabbit anti neun, by Cell Signaling Technology (1 mentions)
Anti fluorescence quencher, by Beyotime (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Rabbit anti app ctf, by Merck Group (1 mentions)
Anti rock1 antibody, by Abcam (1 mentions)
Rabbit anti bace1, by Cell Signaling Technology (1 mentions)
Gapdh, by Merck Group (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Y 27632, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Bx51 fluorescence microscope, by Olympus (1 mentions)
4 protocols using mouse anti aβ
1
Immunohistochemical Analysis of Brain Tissues
2
Dual Immunofluorescence Labeling of Amyloid-β and Neuronal Nuclei
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The frozen brain sections were rehydrated and treated with L.A.B. solution (Polyscience, Inc., Niles, IL, USA) for 20 min. They were then blocked with goat serum for 30 min, and incubated overnight at 4 °C with both the mouse-anti-Aβ (1:400) and rabbit-anti-NeuN (1:400, Cell Signaling Technology, Danvers, MA, USA) antibodies. Subsequently, sections were incubated in DyLight 594-labeled goat anti-mouse IgG and 488-labeled goat anti-rabbit IgG (1:400, Cell Signaling Technology, Danvers, MA, America) at room temperature for 1 h. Finally, the sections were labeled with 4′,6-diamidino-2-phenylindole (DAPI) (Beyotime Institute of Biotechnology, Beijing, China) and sealed with anti-fluorescence quencher (Beyotime Institute of Biotechnology, Beijing, China). The sections were observed and photographed using a confocal fluorescence microscope (Leica, SP8., Wetzlar, Germany).
3
Alzheimer's Disease Pathway Profiling
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Mouse rabbit anti‐APP (Novus, 1:10,000), rabbit anti‐APP‐CTF (Sigma‐Aldrich, 1:5,000), anti‐ROCK1 antibody (Abcam, 1:500), rabbit anti‐SP1 (Abcam, 1:1,000), rabbit anti‐SP6 (Sigma‐Aldrich, 1:1,000), mouse anti‐sAPPα (IBL, 1:50), mouse anti‐Aβ (Cell Signaling, 1:1,000), and rabbit anti‐BACE1 (Cell Signaling, 1:1,000) were used. Loading controls (GAPDH, Sigma‐Aldrich, 1:1,000) were used for Western blot standardization. Lipofectamine 2000 was sourced from Invitrogen. Y‐27632 was purchased from Abcam.
4
Immunofluorescence Staining of Mouse Brain Sections
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IF staining was performed on 3-μm-thick paraffin-embedded mouse brain sections. The sections were deparaffined and boiled in citric acid buffer solution for 5 min. After being cooled to room temperature, the sections were permeabilized in 1% Triton X-100 in PBS for 5 min. Then, the sections were blocked with serum. To detect in situ antigens, the sections were incubated with primary antibodies [mouse anti-Aβ (1:100; catalog number: #15126S; Cell Signaling Technology), rabbit anti-synaptophysin (SYN) (1:100, catalog number: 17785-1-AP; Proteintech, Chicago, IL, USA), and rabbit anti-ionized calcium-binding adapter molecule 1 (IBA-1) (1:100, catalog number: 10904-1-AP; Proteintech)] overnight. Successively, the sections were washed by PBS and incubated with secondary antibodies [Alexa Fluor 594-conjugated goat anti-rabbit IgG (H+L) cross-adsorbed ReadyProbes secondary antibody (catalog number: #R37117) and Alexa Fluor 488-conjugated goat anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody (catalog number: #A-11034) (1:2,000, Invitrogen; Thermo Fisher Scientific, Carlsbad, CA, USA)]. The sections were counterstained with 4′,6-diamidino-2-phenylindole (DAPI) (1 μg/ml; Sigma-Aldrich) and mounted with coverslips, and photos were captured under a BX51 fluorescence microscope (Olympus, Tokyo, Japan).
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