Synapsin 1

Manufactured by Merck Group

Sourced in United States, United Kingdom, Germany

Synapsin 1 is a lab equipment product manufactured by Merck Group. It is a presynaptic protein involved in the regulation of neurotransmitter release and the maintenance of the reserve pool of synaptic vesicles. Synapsin 1 plays a role in the organization and clustering of synaptic vesicles and their attachment to the cytoskeleton.

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Lab products found in correlation

Psd95, by Abcam (4 mentions) Caspase 1, by Santa Cruz Biotechnology (3 mentions) Nf κb p65, by Abcam (3 mentions) Gapdh, by Abcam (3 mentions) Axio observer z1, by Zeiss (3 mentions) Lamin b, by Proteintech (2 mentions) Il 1β, by Santa Cruz Biotechnology (2 mentions) Il 18, by Abcam (2 mentions) Pierce bca protein assay kit, by Beyotime (2 mentions) Gsdmd, by Santa Cruz Biotechnology (2 mentions) Caspase 11, by Santa Cruz Biotechnology (2 mentions) Nlrp3, by Abcam (2 mentions) Gad67, by Merck Group (2 mentions) Nestin, by Merck Group (2 mentions) Glial fibrillary acidic protein (gfap), by Merck Group (2 mentions) Synaptophysin, by Santa Cruz Biotechnology (2 mentions) Slitrk1, by Abcam (2 mentions) Slitrk3, by Abcam (2 mentions) α tubulin, by Merck Group (2 mentions) Glua1, by Santa Cruz Biotechnology (2 mentions)

Close spelling variants of this product

Synapsin (10 citations) Anti-Synapsin-1 (7 citations) Rabbit anti-Synapsin 1 (6 citations) Synapsin-1 antibody (2 citations)

17 protocols using synapsin 1

Quantitative Analysis of Synaptic Markers in Hippocampal Neurons

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Cultured WT or KLHL1-KO hippocampal neurons were fixed at 11 DIV with 4% paraformaldehyde (Electron Microscopy Sciences, Hatfield, PA) following standard methods (Perissinotti et al., 2014 (link)). Primary antibodies were diluted in blocking solution containing 2% goat serum (Jackson ImmunoResearch Laboratories, West Grove, PA) plus 0.4% saponin in PBS and incubated overnight at 4°C (MAP2, 1:1000, EnCor Biotechnology, FL; synapsin I, 1:1000, Millipore, CA; gephyrin, 1:500, SySy, Germany; GAD67 1:500, Thermo Scientific; PSD-95, 1:1000, NeuroMab; VGlut-1, 1:1000, SySy; synapsin I, 1:1000, Millipore, CA). Samples were incubated in alexa fluor-conjugated secondary antibodies (1:2,000; Life Technologies) for 1 h at room temperature. Coverslips were mounted on slides with Citiflour (Ted Pella, Redding, CA) and stored at −20°C for subsequent detection (Multiphoton Leica TCS SP5). Data was analyzed with ImageJ freeware (NIH) (Rasband, 1997–2006 ). Images were thresholded using the Otsu plugin. Synaptic puncta number was measured in n sample areas = 246 μm² each. The JACoP plugin was used to calculate Mander’s correlation coefficients (co-localization percentages), which provides a good quantification of signal co-localization between samples of different intensities (Otsu, 1979 (link); Bolte and Cordelières, 2006 (link)).

Perissinotti P.P., Ethington E.A., Almazan E., Martínez-Hernández E., Kalil J., Koob M.D, & Piedras-Rentería E.S. (2015). Calcium current homeostasis and synaptic deficits in hippocampal neurons from Kelch-like 1 knockout mice. Frontiers in Cellular Neuroscience, 8, 444.

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Reagents and Antibodies for Neuronal Studies

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The CMV-mCherry-dynamitin expression vector was kindly shared by M. Meffert (Johns Hopkins, MD; Shrum et al., 2009 (link)) while the mCherry plasmid was a gift from R.Y. Tsien (UC San Diego, CA). The 4xGFP construct was a gift from W. Hampe (UMC Hamburg-Eppendorf, Hambug; Seibel et al., 2007 (link)). Commercial plasmids include Dendra2 (Evrogen) and CRTC1 (Open Biosystems, Huntsville, AL). Antibodies used in all these experiments include: rabbit polyclonal antibodies against CRTC1 (Bethyl, Montgomery, TX and Proteintech, Chicago, IL), pCRTC1(S151; Bethyl) Dendra2 (Evrogen, Moscow, Russia), TUJ1 (Covance, Princeton, NJ), Dynein heavy chain (Santa Cruz, Dallas, TX), and phosphoCREB-S133 (Cell Signaling); mouse monoclonal antibodies against PSD95 (Thermoscientific, Rockford, IL), synapsin1 (Millipore, Billerica, MA), CamKIIα (Millipore), HA-epitope (Sigma), GAPDH (Fitzgerald, Acton, MA), GFP (Clontech, Mt. View, CA), GAD67 (Millipore), and KPNB1 (ABR, Golden, CO); polyclonal chicken antibody against MAP2 (Phosphosolutions, Aurora, CO) and synaptotagmin (Chemicon, Temecula, CA). All secondary antibodies are conjugated to Alexa dyes (488, 546, 555, 568, and 633; Invitrogen).

Ch'ng T.H., DeSalvo M., Lin P., Vashisht A., Wohlschlegel J.A, & Martin K.C. (2015). Cell biological mechanisms of activity-dependent synapse to nucleus translocation of CRTC1 in neurons. Frontiers in Molecular Neuroscience, 8, 48.

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Quantitative Protein Analysis via Western Blotting

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Protein quantification was performed with a Pierce BCA Protein Assay Kit (Beyotime Institute of Biotechnology), and 30–50 μg of total protein was resolved by polyacrylamide gel electrophoresis (SDS-PAGE, 8%). Protein levels were determined via incubation with antibodies against NF-κB-p65 (1:500; Abcam, UK), IκBα (1:500; Abcam, UK), NLRP3 (1:500; Abcam, UK), caspase-1 (1:500; Santa Cruz Biotechnology, USA), caspase-11 (1:500; Santa Cruz Biotechnology, USA), IL-1β (1:500; Santa Cruz Biotechnology, USA), IL-18 (1:500; Abcam, UK), Synapsin-1 (1:500; Millipore, USA), PSD-95 (1:500; Abcam, UK), GSDMD (1:500; Santa Cruz Biotechnology, USA), Lamin B (1:1000; Proteintech, USA), and GAPDH (1:500; Abcam, UK). The blots were imaged with ECL Plus western blotting detection reagents. ImageJ software was used to determine the average absorbance value of the corresponding bands.

Dai J., Li X., Wang C., Gu S., Dai L., Zhang J., Fan Y, & Wu J. (2021). Repeated neonatal sevoflurane induced neurocognitive impairment through NF-κB-mediated pyroptosis. Journal of Neuroinflammation, 18, 180.

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Immunohistochemical Characterization of Neural Tissue

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Slides were thawed to room temperature, and lines drawn to partition the edges of sections on the glass slides using a PAPpen (ImmEdge Pen; Vector Labs H4000). The sections were rehydrated, blocked, permeabilized and immunostained with primary antibodies: PAX6 (BioLegend, 1:200), SOX2 (Santa Cruz, 1:100), β-tubulin III (Sigma, 1:1000), FOXG1 (Takara, 1:500), SOX10 (Santa Cruz, 1:100), Ki67 (Thermo Fisher, 1:200), Nestin (Millipore, 1:200), GFAP (Millipore, 1:400), S100beta (DAKO Potts, 1:400), CTIP2 (Abcam, 1:500), SATB2 (Abcam, 1:50), MAP2AB (Abcam, 1:1000-2000), NeuN (Millipore, 1:200), TBR1(Abcam, 1:500), BRN2 (Millipore, 1:500), MEF2C (Novis, 1:200), AT8 (Invitrogen, 1:1000), PHF1 (gift from Peter Davies,1:1000), Total Tau (DAKO Potts, 1:200), DA9 (gift from Peter Davies, 1:1000), ELAVL4 (Santa Cruz, 1:200), TIA1 (Santa Cruz, 1:100), G3BP1 (Protein Tech, 1:200), VGLUT1 (gift from Susan Morton, Tom Jessell, 1:16,000), Calbindin1 (Swant, 1:10,000), Homer1 (SYSY, 1:250) and Synapsin1 (Millipore, 1:100). Primary antibodies were incubated overnight at 4C, washed three times with PBS and then incubated with corresponding Alexa Fluor conjugated secondary antibody (1:333-1000) for 1 hour at room temperature. Sections were coverslipped and imaged using fluorescence and confocal microscopy (Zeiss AXIO Observer.Z1; Zeiss 780).

Bowles K.R., Silva M.C., Whitney K., Bertucci T., Berlind J.E., Lai J.D., Garza J.C., Boles N.C., Mahali S., Strang K.H., Marsh J.A., Chen C., Pugh D.A., Liu Y., Gordon R.E., Goderie S.K., Chowdhury R., Lotz S., Lane K., Crary J.F., Haggarty S.J., Karch C.M., Ichida J.K., Goate A.M, & Temple S. (2021). ELAVL4, splicing, and glutamatergic dysfunction precede neuron loss in MAPT mutation cerebral organoids. Cell, 184(17), 4547-4563.e17.

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Western Blot Analysis of Mitochondrial Proteins

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Proteins (40 μg per lane) were separated with SDS-PAGE and then transferred to a polyvinylidene fluoride membrane (Millipore, Billerica, MA, USA). After being blocked with 5% skim milk in TBST for 1 h, the membranes were incubated with primary antibody against Dynamin-related protein 1 (DRP1; 1:1,000; Abcam), Mitofusin 2 (MFN2; 1:1,000; Abcam), NLRP3 (1:1,000; Cell Signaling Technology, Danvers, MA, USA), caspase-1 (1:1,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), GSDMD (1:500; Abcam), interleukin-1β (IL-1β; 1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA), IL-18 (1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA), synapsin 1 (1:2,500; Millipore, Billerica, MA, USA), PSD-95 (1:1,500; Abcam), voltage-dependent anion channel (VDAC; 1:1,000; Cell Signaling Technology, Danvers, MA, USA), or GAPDH (1:1,000; Cell Signaling Technology, Danvers, MA, USA). After three washes, the membranes were treated with species-specific peroxidase-conjugated secondary antibodies for 1 h at room temperature. Bands were visualized by enhanced chemiluminescence and quantified with Image Quant Software (Syngene).

Zuo Y., Yin L., Cheng X., Li J., Wu H., Liu X., Gu E, & Wu J. (2020). Elamipretide Attenuates Pyroptosis and Perioperative Neurocognitive Disorders in Aged Mice. Frontiers in Cellular Neuroscience, 14, 251.

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Comprehensive Antibody Panel for Neuroscience

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Primary antibodies used for WB and Immunocytochemistry: ISL1/2 (DSHB, 39.4D5-c), CHAT (Millipore, AB144P), ITGB1 (Millipore, clone HUTS-4, MAB2079Z), ITGB4 (Millipore, MAB1964), ILK (Cell Signaling, 3862), p-FAK (Cell Signaling, 3281S), FAK (Cell Signaling, 3285S), ACTIN (Sigma Aldrich, A2066), KI67 (Abcam, ab66155), MAP2 (Biolegend, 840601), TUJ1 (Biolegend, 802001), PSD95 (NeuroMab, 75–028), SYNAPTOPHYSIN (Abcam, ab32127), SYNAPSIN-1 (Millipore, AB1543), FOXA-2 (Santa Cruz Biotechnology, sc-101060), NeuN (Biolegend, 834502), Laminin alpha-1 (Santa Cruz, sc-74417), GFP (Abcam, ab6673), SOD1 (Proteintech, 10269–1-AP), Ubiquitin (Santa Cruz Biotechnology, sc-8017) and GABA (Sigma Aldrich, A2052).

Álvarez Z., Ortega J.A., Sato K., Sasselli I.R., Kolberg-Edelbrock A.N., Qiu R., Marshall K.A., Nguyen T.P., Smith C.S., Quinlan K.A., Papakis V., Syrgiannis Z., Sather N.A., Musumeci C., Engel E., Stupp S.I, & Kiskinis E. (2023). Artificial Extracellular Matrix Scaffolds of Mobile Molecules Enhance Maturation of Human Stem Cell-Derived Neurons. Cell stem cell.

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Quantifying Protein Levels with Western Blot

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Protein quanti cation was performed using the Pierce BCA Protein Assay Kit (Beyotime Institute of Biotechnology), and 30-50 μg of total protein was dissolved by polyacrylamide gel electrophoresis (SDS-PAGE, 8%). Protein levels were determined via incubation with antibodies against NF-κB-p65 (1:500; Abcam, UK), IκBα (1:500; Abcam, UK), NLRP3 (1:500; Abcam, UK), caspase-1 (1:500; Santa Cruz Biotechnology, USA), caspase-11 (1:500; Santa Cruz Biotechnology, USA), IL-1β (1:500; Santa Cruz Biotechnology, USA), IL-18 (1:500; Abcam, UK), synapsin 1 (1:500; Millipore, USA), PSD-95 (1:500; Abcam, UK) GSDMD (1:500; Santa Cruz Biotechnology, USA), Lamin B (1:1000; Proteintech, USA), and GAPDH (1:500; Abcam, UK). The blots were imaged using ECL Plus western blotting detection reagents. Image J software was used to determine the average absorbance value of the corresponding bands.

Dai J., Li X., Wang C., Gu S., Dai L., Zhang J., Fan Y., & Wu J. (2021). Repeated Neonatal Sevoflurane Induced Neurocognitive Impairment Through NF-κB-Mediated Pyroptosis.

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Generation and Characterization of Antibodies

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SALM3 (1929) guinea pig polyclonal antibodies were generated using a synthetic peptide as immunogen (aa 594–608 of mouse SALM3; CYGYARRLGGAWARR). Peptides mimicking the last 30 aa of mouse SALM1, SALM2, and SALM4 were used to generate guinea pig polyclonal antibodies (2022, 2058, and 2026, respectively). The following antibodies have been described: PSD-95 (1690) (Han et al., 2010 (link)), NGL-3 (2020) (Lee et al., 2014 (link)), SALM3 (1816), and SALM5 (1907) (Mah et al., 2010 (link)). The following antibodies were purchased: α-tubulin, synapsin I (Sigma), synaptophysin, GluA1, GluA2 (Santa Cruz), slitrk1, slitrk3 (Abcam), GluN1 (Invitrogen), GluN2A (Zymed), GluN2B (NeuroMab), PTPσ (17G7.2, mouse, Medimabs), and IL-1RAcP (Millipore). IL1RAPL1 antibody was a kind gift from Dr. Carlo Sala.

Li Y., Zhang P., Choi T.Y., Park S.K., Park H., Lee E.J., Lee D., Roh J.D., Mah W., Kim R., Kim Y., Kwon H., Bae Y.C., Choi S.Y., Craig A.M, & Kim E. (2015). Splicing-Dependent Trans-synaptic SALM3–LAR-RPTP Interactions Regulate Excitatory Synapse Development and Locomotion. Cell reports, 12(10), 1618-1630.

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Quantitative Western Blot Analysis of Cortical Proteins

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For Reelin protein levels, protein extracts were prepared from cortical tissue extracted from WT and KO P2 mice. For other proteins, extracts were prepared from somatosensory cortical tissue, dissected from 400-μm-thick brain sections of WT and KO adult mice. Western blotting was performed using 10–12% SDS polyacrylamide precast gels (BIO-RAD Laboratories) with 15–30 μg of protein extracts loaded per lane. Protein transfer on nitrocellulose membrane was done by using the Trans-Blot Turbo Transfer System (BIO-RAD Laboratories). Nitrocellulose filters were incubated with primary antibodies raised against Reelin (G10, 1:1000; GeneTex), NeuN (A60, 1:1000; Chemicon), GFAP (G-A-5, 1:500; Sigma-Aldrich), GAD65 (GAD-6, 1:1000; abcam), GAD67 (1G10.2, 1:1000; Merck Millipore), Synapsin I (S193, 1:2000; Sigma-Aldrich), and GAPDH (6C5, 1:500; Merck Millipore) overnight at 4°C, followed by IRDye 800CW- or 680LT-conjugated secondary antibodies (1:1500; LI-COR). To visualize bands, Odyssey Fc Imager was used. Densitometric analysis was performed using Image Studio Software (LI-COR), and each band was normalized to the GAPDH signal in each lane and expressed as percentage.

Bonini S.A., Mastinu A., Maccarinelli G., Mitola S., Premoli M., La Rosa L.R., Ferrari-Toninelli G., Grilli M, & Memo M. (2016). Cortical Structure Alterations and Social Behavior Impairment in p50-Deficient Mice. Cerebral Cortex (New York, NY), 26(6), 2832-2849.

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Generation and Characterization of Antibodies

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