The levels of molecules or enzymes involving Aβ metabolism, phosphorylated Tau, and synapse-related proteins were analyzed using Western blotting. Proteins in the animal brain homogenate were extracted with RIPA buffer. Samples were loaded on SDS-PAGE (4–10 % acrylamide) gels. Separated proteins were transferred to nitrocellulose membranes. The blots were probed with the following antibodies: anti-APP C-Terminal (171610, Millipore) which recognizes full-length APP (APPfl) and C-terminal fragment (CTF)-β, anti-BACE1 (Millipore), anti-NEP (Millipore), anti-receptor for advanced glycosylation products (RAGE, Millipore), anti-LRP-1 (5A6, Calbiochem), anti-IDE (Epitomics), anti-phosphorylated-Tau antibodies including anti-pS396 (Signalway) and anti-pS199 (epitomics), anti-Synaptophysin (Millipore), anti-Synapsin-1 (Millipore), anti-PSD95 (Millipore), anti-PSD93 (Millipore) and anti-β-actin (Sigma-Aldrich). The membranes were incubated with IRDye 800CW secondary antibodies (Li-COR) and scanned using the Odyssey fluorescent scanner. The band density was normalized to β-actin for analysis.
Anti synapsin 1
Manufactured by Merck Group
Sourced in United States
Anti-Synapsin-1 is a primary antibody used in immunohistochemistry and Western blotting applications. It specifically targets the Synapsin-1 protein, which is a presynaptic phosphoprotein involved in the regulation of neurotransmitter release.
Automatically generated - may contain errors
Lab products found in correlation
Anti synaptophysin, by Merck Group (1 mentions)
Anti bace1, by Merck Group (1 mentions)
Anti psd95, by Merck Group (1 mentions)
Irdye 800cw secondary antibody, by LI COR (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Psd95, by Cell Signaling Technology (1 mentions)
P creb, by Cell Signaling Technology (1 mentions)
Brain derived neurotrophic factor (bdnf), by Santa Cruz Biotechnology (1 mentions)
Glun2a, by Abcam (1 mentions)
Glur2, by Merck Group (1 mentions)
Glur1, by Merck Group (1 mentions)
Glun2b, by Abcam (1 mentions)
Anti synaptotagmin1, by Abcam (1 mentions)
Histone 3, by Abcam (1 mentions)
Acetylated h3, by Merck Group (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
β actin, by Abcam (1 mentions)
Mouse anti oct3 4, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti pax6, by BioLegend (1 mentions)
Rabbit anti nanog, by ReproCELL (1 mentions)
7 protocols using anti synapsin 1
1
Quantifying Brain Protein Levels in AD
2
Hippocampal Protein Expression Analysis
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Western blotting was performed according to methods established in our laboratory46 (link). Hippocampus was removed and homogenized on ice with RIPA buffer (Bi Yuntian). The samples were separated by SDS-PAGE, transferred onto nitrocellulose membranes and then covered by 5% non-fat milk. After washings by Tween-TBS, the membranes were detected by primary antibodies overnight, and then by secondary antibodies conjugated to IRDyeTM (800CW) for 1 h. The visual blots were acquired from the Odyssey Infrared Imaging System (Licor biosciences, Lincoln, NE, USA). The full length blots are presented in Supplementary Information. The primary antibodies are displayed as follows: CREB (cell signaling, 1:1000), p-CREB (cell signaling, 1:1000), BDNF (SANTA CRUZ, 1:1000), GluN2A (abcam, 1:1000), GluN2B (abcam, 1:1000), GluR1 (Millipore, 1:500), GluR2 (Millipore, 1:500), PSD95 (cell signaling, 1:1000), anti-Synaptotagmin1 (abcam, 1:2000), anti-Synapsin1 (Millipore, 1:1000), PT231 (SAB, 1:500), PP1 (Millipore, 1:200), HT7 (Thermo, 1:500), β-actin (abcam, 1:1000), acetylated H3 (Millipore, 1:1000), acetylated H3K14 (Millipore, 1:1000), acetylated H3K9 (Millipore, 1:1000), Histone 3 (abcam, 1:1000).
3
Immunofluorescence Staining of Neural Markers
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Immunofluorescence staining was conducted as reported previously.10 ,25 (link),26 (link),28 (link),29 (link) The following antibodies were used: rabbit anti-neurofilament middle chain (Millipore, Billerica, MA, USA), mouse anti-βIII tubulin (Promega, Madison, WI, USA), rabbit anti-CRIM1 (Sigma-Aldrich), rat anti-CTIP2 (Abcam, Cambridge, MA, USA), anti-Fezf2 (Abcam), rabbit anti-Foxp2 (Abcam), mouse anti-human Neural cell adhesion molecule (NCAM) (hNCAM; Santa Cruz Biotechnology, Dallas, TX, USA), antisynapsin1 (Millipore), rabbit anti-Nanog (ReproCELL, Kanagawa, Japan), mouse anti-Oct3/4 (Santa Cruz Biotechnology), rabbit anti-Pax6 (BioLegend, San Diego, CA, USA), and mouse anti-human nuclei (Abnova, Taipei City, Taiwan).
4
Synapsin I Expression in TBI Mice
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The total proteins from cortical tissue of TBI-affected hemisphere in sham-operated, sTBI and mTBI mice were extracted, and Synapsin I was analyzed by Western blot. Briefly, protein samples were separated by electrophoresis on a 10% polyacrylamide gel and electrotransferred to a nitrocellulose membrane. Nonspecific binding sites were blocked in TBS, overnight at 4 °C, with 2% BSA and 0.1% Tween-20. Membranes were rinsed for 10 min in a buffer (0.1% Tween-20 in TBS) and then incubated with anti-synapsin I (1:1500, Sigma) followed by anti-rabbit IgG horseradish peroxidase conjugate (Santa Cruz Biotechnology). After rinsing with buffer, the immunocomplexes were developed with G:BOX Chemi XX6 gel doc system (Syngene) and analyzed in Image Lab software. β-Actin was used as an internal control for Western blot.
5
Neuronal Marker Detection in Transdifferentiated Cells
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Neuronal markers were detected after treatments under immunofluorescence microscope (Olympus, Japan) at 40X objective as describe previously (Naveen et al., 2016) . Briefly, transdifferentiated cells were fixed with 2% paraformaldehyde and washed with PBS. Cells were permibialised using 1.5% tween in PBS. Blocking was done with 2%FBS in PBS after washing thrice. Anti-Map2 a,b,c (MA5-12826; Thermo Fisher Scinetific;1:500), anti-Neuron Specific Enolase (NSE; PA5-12374; Thermo Fisher Scientific; 1:500), anti-nestin (MA1-110; Thermo Fisher Scientific; 1:500), anti-nurr1 (PA5-22799; Thermo Fisher Scientific; 1:500), anti-synapsin I (S193; Sigma;1:1000), anti-Prominin (SAB4300882; Sigma-Aldrich; 1:1000) (Sigma Aldrich, USA), anti-N-Cadherin (C3865; Sigma;1:500), anti-glial fibrillary acidic protein (GFAP; 14-9892; eBioscience, 1:1000) FITC-Labelled anti-CD11c (553801; BD biosciences; 1:500) were used as primary antibodies. Secondary antibodies like goat antimouse IgG/IgM Alexa flour 488/543 (A11029; Thermo Fisher Scinetific; 1:2000), goat anti-Rabbit Alexa Flour 546 (A11003; Thermo Fisher Scinetific; 1:2000) were used. All the images
6
Investigating HIV-1 Protein Interactions
7
Protein Expression and Immunoblotting Analysis
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Proteins were loaded and separated on a 12% Bis-Tris polyacrylamide gel (NuPAGE, Invitrogen) and processed as described previously (Zhu et al., 2015a) . Primary antibodies anti-PrP antibody POM1 (400 ng mL À1 ), anti-NeuN clone EPR12763 (1:3000, Abcam, ab177487), anti-synaptophysin clone 2/synaptophysin (1:10,000, BD Biosciences, 611880), anti-synapsin I (1:2000, Millipore, AG, USA, AB1543), anti-Iba1 antibody (1:1000; Wako Chemicals GmbH, Germany, 019e19,741), anti-GFAP antibody (D1F4Q) XP Rabbit mAb (1:3000; Cell Signaling Technology, 12,389), anti-Mfge8 antibody (1:1000; R&D Systems, AF2805), anti-actin antibody (1:10,000, Millipore), and secondary antibody horseradish peroxidaseeconjugated goat anti-mouse IgG (1:10,000, Jackson ImmunoResearch, 115-035-003), goat anti-rabbit IgG (1:10,000, Jackson ImmunoResearch, 111-035-045), and donkey anti-goat IgG(1:10,000, Jackson ImmunoResearch, 705-035-147) were used.
For proteinase K (PK) digestion, samples were adjusted to 20 mg protein in 20 mL and digested with 25 mgm L À1 proteinase K for 30 minutes at 37 C.
For proteinase K (PK) digestion, samples were adjusted to 20 mg protein in 20 mL and digested with 25 mgm L À1 proteinase K for 30 minutes at 37 C.
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