The Aβ1-42 used in these experiments was acquired from Life Technologies (USA); β-asarone was purchased from NIFDC (Beijing, China), and the purity value is 96.8%; donepezil was obtained from the First Affiliated Hospital of Jinan University (Guangzhou, China); A3 was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); CSA was purchased from Selleck (Houston, Texas, USA); 3MA, high-glucose DMEM, FBS, trypsin, and PBS were obtained from Gibco (Gaithersburg, MD, USA); Cell Counting Kit-8 (CCK-8), Cytotoxicity LDH Assay Kit—WST (LDH), and Cellular Senescence Detection Kit—SPiDER-βGal were obtained from Dojindo Molecular Technologies, Inc. (Tokyo, Japan); the bicinchoninic acid (BCA) protein assay kit, Immunol Fluorescence Staining Kit, and Immunohistochemistry Staining Kit were obtained from Beyotime (Shanghai, China); anti-SQSTM1/P62, anti-BECN, anti-LC3 II, anti-beta amyloid, anti-BACE1, anti-synapsin1, anti-Parkin, anti-Pink1, anti-APPL, and anti-PS1 were obtained from Abcam (Cambridge, UK).
Anti synapsin 1
Manufactured by Abcam
Sourced in United States, Germany
Anti-synapsin 1 is an antibody that binds to the synapsin 1 protein. Synapsin 1 is a neuronal phosphoprotein that plays a role in the regulation of neurotransmitter release and synaptic vesicle function.
Automatically generated - may contain errors
Lab products found in correlation
Anti psd95, by Abcam (2 mentions)
High glucose dmem, by Thermo Fisher Scientific (1 mentions)
Anti sqstm1 p62, by Abcam (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Aβ1 42, by Thermo Fisher Scientific (1 mentions)
Anti parkin, by Abcam (1 mentions)
Donepezil, by Santa Cruz Biotechnology (1 mentions)
Bicinchoninic acid bca protein assay kit, by Beyotime (1 mentions)
Anti pink1, by Abcam (1 mentions)
Anti lc3ii, by Abcam (1 mentions)
Immunol fluorescence staining kit, by Beyotime (1 mentions)
Anti ps1, by Abcam (1 mentions)
Anti beta amyloid, by Abcam (1 mentions)
Anti bace1, by Abcam (1 mentions)
Fitc dextran, by Merck Group (1 mentions)
Goat anti mouse igg, by Santa Cruz Biotechnology (1 mentions)
Autosampler vial, by Thermo Fisher Scientific (1 mentions)
Anti cox 2, by Abcam (1 mentions)
12 protocols using anti synapsin 1
1
Alzheimer's Disease Pathogenesis Model
2
Biochemical Markers Quantification Protocol
3
Reverse Co-Immunoprecipitation of Synaptic Proteins
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For reverse co-IPs, protein G-Sepharose 4 Fast Flow beads (GE Healthcare) were pre-bound with 4 μg of the following antibodies, respectively: anti-mitochondrial creatine kinase U-type (MtCK, Abcam, San Francisco, catalog no. ab76506); anti-β-synuclein (Abcam, catalog no. ab76111); anti-apoE (Abcam, catalog no. ab1906); anti-apoAI (Santa Cruz Biotechnology, Dallas, TX, catalog no. sc-30089); anti-syntaxin 1B (Synaptic Systems, Göttingen, Germany, catalog no. 110402); anti-synapsin 1 (Abcam, catalog no. ab8); anti-synaptogyrin-3 (Santa Cruz Biotechnology, catalog no. sc-68936); anti-synaptophysin (Santa Cruz Biotechnology, catalog no. sc-9116); and anti-synaptotagmin (Millipore, catalog no. MAB5200). These samples were then incubated with 1 mg of lysate at 4 °C overnight. To verify the binding preference of Tau isoforms, dephosphorylated lysates were used adapting a dephosphorylation protocol obtained from New England Biolabs. Beads were washed extensively with IP washing buffer and eluted in 2× SDS-PAGE sample buffer for subsequent Western blot analysis. Blots were probed with the following antibodies: anti-creatine kinase U-type; mitochondrial (MtCK); anti-β-synuclein; anti-apoE; anti-apoAI; anti-syntaxin 1B; anti-synapsin 1; anti-synaptogyrin-3; anti-synaptophysin; anti-synaptotagmin, and anti-Tau. For quantification, ImageJ was used.
4
Quantifying Synaptic Protein Expression
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The mouse NPS (SFRNGVGSGAKKTSFRRAKQ) was purchased from Cellmano Biotech Ltd. (Hefei, China), and dissolved in saline to prepare stock solutions. Antibodies were purchased from several companies: Anti-c-Fos (Cat. No. sc-166940) from Santa Cruz (Santa Cruz, CA, USA); Anti-NPSR (Cat. No. ab92425), Anti-DM1A (Cat. No. ab7291) and Anti-PS1 (Cat. No. ab76083) from Abcam (Cambridge, MA, USA); Anti-synapsin-1 (Cat. No. 2908691), Anti-APP (Cat. No.2986158) from Millipore (Billerica, MA, USA); Anti-Phospho-APP (Thr668; Cat. No. P05067), Anti-postsynaptic density protein 95 (PSD95; Cat. No. P78352) from Cell Signaling (MA, USA); Anti-4G8 (Cat. No. 800701) from BioLegend (USA); and Rhodamine Red-X-conjugated goat anti-rabbit/mouse IgG (Cat. No. BA1031; BA1032) from Boster Biological Technology (Wuhan, China). The BCA kit (Cat. No. 23227) was purchased from Beyotime Biotechnology (Shanghai, China).
5
Quantitative Western Blot Analysis of AD Markers
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Brain samples were homogenized in ice‐cold RIPA lysis buffer. Protein samples were loaded on a 4%–20% SDS‐polyacrylamide gel and then transferred onto nitrocellulose (NC) membranes. After blocking with 5% fat‐free milk, the NC membranes were incubated with the following primary antibodies overnight at 4°C: anti‐APP C‐terminal (1:1000, Biolegend); anti‐6E10 (1:400, Biolegend); anti‐RAGE (1:1000, Millipore); anti‐LRP‐1 (1:1000, Abcam); anti‐pS396‐tau (1:1000, Abcam); anti‐pT231‐tau (1:1000, Signalway); anti‐Tau5 (1:1000, Millipore); anti‐Synapsin‐1 (1:1000, Abcam); anti‐PSD95 (1:1000, Millipore); and anti‐β‐actin (1:2000, Origene). The membranes were incubated with the corresponding IRDye 800 CW‐conjugated secondary antibodies and scanned using an Odyssey fluorescent scanner. Relative band intensities were normalized to the band intensity of the internal reference protein for analysis.
6
Immunofluorescence Staining of Cultured Cells
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Cells were washed with PBS and then fixed with 4% paraformaldehyde (with 4% sucrose) for 15 min at room temperature. The cells were washed twice (5 min each wash) with Tris-buffered saline (TBS) and permeablized with 0.1% Triton X-100 in TBS (TBS-Tx) for 5 min. They were blocked with 5% donkey serum in TBS-Tx for 2 h at room temperature. They were then incubated with the primary antibodies overnight at 4 °C. Primary antibodies used were: anti-GFP (1:3000, Rockland), anti-MAP2 (1:1000, Sigma), and anti-Synapsin1 (1:500, Abcam). After three washes with TBS-Tx, the cells were incubated with secondary Alexa Fluor® antibodies (1:500, Thermo Fisher Scientific) for 2 h at room temperature. The cells were stained with DAPI (1:5000) for 10 min at room temperature before they were mounted onto glass microscope slides and allowed to dry before imaging.
7
Glycan-Specific Antibody Characterization
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The following antibodies were used: Anti-Paucimannose (serum-free undiluted cell culture supernatant; clone: Mannitou, gift from B. Schmitz, Bonn, Germany); anti high-mannose (serum-free undiluted cell culture supernatant; clone 492, gift from B. Schmitz, Bonn, Germany); anti-polySia (1:10,000 dilution, clone 735 gift from R.Gerady-Schahn, Hannover Germany, IgG), anti-NCAM (1:1000 dilution clone 5B8 IgG Hybridoma Bank); anti-CML (1:10,000 dilution; clone CML56 abcam); anti-O-GlcNAc (1:10,000 dilution; clone CTD110.6, Cell Signaling); anti-Synapsin-1 (1:5000 dilution; polyclonal 64581 Abcam); anti-RAGE (1:1000 dilution; polyclonal 3611 Abcam).
8
Western Blot Analysis of Synaptic Proteins
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Homogenized slice lysates were prepared with lysis buffer (PRO-PREPTM, Intron Biotechnology, Burlington, MA, USA). Proteins were resolved by SDS-PAGE (100 µg/lane) using 10% (w/v) polyacrylamide gels and transferred to PVDF membranes (Millipore). Membranes were incubated with antibodies against anti-PSD 95 (1:5000, Abcam, Cambridge, MA, USA); anti-synapsin I (1:3000, Abcam); superoxide dismutase (SOD) (1:7500, Abcam); and Nrf2 (1:5000, Abcam) for 2 h at room temperature. As a loading control, membranes were also probed with anti-β-actin antibody (1:10,000, Abcam). The reaction was developed with an enhanced chemiluminescence Western blot analysis system (ECL, GE Healthcare, Marlborough, MA, USA). Signal intensities were analyzed using a gel-scanning integrated optical density software program (Multi-gauge 3.0, Fuji film, Tokyo, Japan).
9
Neuronal ER Stress Modulation Assay
10
Immunofluorescence Staining of Stem Cells
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Cells were fixed with 3.7% formaldehyde in phosphate-buffered saline (PBS). After permeablisation with 0.1% to 0.2% Triton X-100/PBS, cells were incubated with a primary antibody, followed by staining with a secondary antibody conjugated with AlexaFluor488 (1:500) or AlexaFluor555 (1:500). The following primary antibodies were used in this study: anti-SSEA4 (1:250; Merck Millipore, Darmstadt, Germany), anti-OCT4 (1:400; Abcam, Cambridge, UK), anti-NANOG (1:20; R&D Systems, Minneapolis, MN, USA), anti-TRA-1-60 (1:250; e-Bioscience, Santa Clara, CA, USA), anti-β-III Tubulin (1:1,000; BioLegend, San Diego, CA, USA), anti-SOX17 (1:200; Abcam), anti-smooth muscle actin (1:250; Dako, Santa Clara, CA, USA), anti-Desmin (1:200; Thermo Fisher Scientific), anti-MAP2 (1:500; Santa Cruz, Dallas, TX, USA), and anti-Synapsin I (1:500; Abcam). Nuclei were counterstained with DAPI using VECTASHIELD mounting medium with DAPI (Vector Laboratories, Burlingame, CA, USA).
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