Snap25

Manufactured by Santa Cruz Biotechnology

Sourced in United States

SNAP25 is a laboratory product used for research purposes. It is a protein that plays a core role in the regulation of neurotransmitter release in neuronal cells. The product is intended for use in scientific research and experimentation, and its function is to facilitate the study of synaptic transmission and related neurological processes.

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Lab products found in correlation

8 protocols using snap25

Comprehensive Molecular Profiling of Neurodegeneration

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The following primary antibodies were used: TH and NeuN (Merck-Millipore, MA, USA); Bcl-2, Bax, Cytochrome c, Caspase-3, PARP-1, phospho-JNK, JNK, Nurr1, DAT, GFAP, Iba-1, PSD-95, SNAP-25, Synaptophysin (SYP), phospho-mTOR (296. Ser2481) and β-actin (Santa Cruz, CA, USA); AMPKα, phospho-AMPKα, phospho-p44/42 MAPK (ERK1/2, Thr202/Tyr204), p44/42 MAPK (ERK1/2), phospho-p38 MAP kinase (Thr180/Tyr182), p38 MAPK, α-synuclein, phospho-CREB (Ser133), CREB, and mTOR (Cell Signaling, MA, USA); VMAT2 and phospho-α-synuclein (Ser129) (Abcam, Cambridge, UK); and phospho-α-synuclein (Ser129, BioLegend, CA, USA). Detailed antibody information is provided in Additional file 1: Table S1.

Park J.S., Choe K., Lee H.J., Park T.J, & Kim M.O. (2023). Neuroprotective effects of osmotin in Parkinson’s disease-associated pathology via the AdipoR1/MAPK/AMPK/mTOR signaling pathways. Journal of Biomedical Science, 30, 66.

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Comprehensive Antibody Panel for Neurodegenerative Markers

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Mouse monoclonal antibody against human CD171 (IgG2a, Thermo Fisher Scientific, Waltham, MA), CD9 (Bio-Rad, Hercules, CA), CD81 (BD Biosciences, Franklin Lakes, NJ), control mouse IgG2a (BioLegend, San Diego, CA), normal mouse IgG (Equitech-Bio), glyceraldehyde-3-phosphate dehydrogenase (GAPDH, BD), aldolase (ALDO, Santa Cruz Biotechnology, Dallas, TX), surfactant protein B (SFTPB, US Biologicals, Salem, MA), presenilin 1 (PSEN, Santa Cruz), β-secretase (BACE, Santa Cruz), S100B (R&D Systems, Minneapolis, MN), acetylcholinesterase (AchE, Santa Cruz), neurogranin (NRGN, BioLegend), cathepsin D (CTSD, Santa Cruz), synaptic vesicle glycoprotein 2A (SV2A, Abcam, Cambridge, United Kingdom), synaptopodin (SYNPO, Santa Cruz), and synaptosome associated protein 25 (SNAP25, Santa Cruz), and enolase 2 (ENO2, R&D) were purchased from designated suppliers.

Hotta N., Tadokoro T., Henry J., Koga D., Kawata K., Ishida H., Oguma Y., Hirata A., Mitsuhashi M, & Yoshitani K. (2022). Monitoring of Post-Brain Injuries By Measuring Plasma Levels of Neuron-Derived Extracellular Vesicles. Biomarker Insights, 17, 11772719221128145.

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Protein Separation and Detection

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The subfractions or purified fractions were mixed with an aliquot of Laemmli buffer (0.125 M Tris–HCl [pH 6.8], 4% sodium dodecyl sulfate [SDS], 20% glycerol, 10 mM mercaptoethanol, and 0.002% bromophenol blue). After boiling for 5 min, the samples were cooled to room temperature, separated using a 15% SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and transferred to a nitrocellulose membrane (Schleicher & Schuell, USA) at 30 mA constant current. The membrane was blocked for 1 h at room temperature with 5% nonfat milk in Tris-buffered saline (20 mM Tris–HCl [pH 7.4], 137 mM NaCl, and 2.7 mM KCl). The blocked nitrocellulose membranes were incubated with rabbit antibodies against nSMase2, SNAP25 (diluted 1:2,000; Santa Cruz Biotechnology), Hsp60 (diluted 1:2,500; Stressgen), and cytochrome c (diluted 1:1,000; BD Biosciences) at 4°C with constant shaking. Antibody-binding sites were detected using an alkaline phosphatase-conjugated IgG antibody (Santa Cruz Biotechnology) and a chromogenic substrate (1-Step^TM NBT/BCIP; Pierce, USA).

Choi J.M., Piao Y., Ahn K.H., Kim S.K., Won J.H., Lee J.H., Jang J.M., Shin I.C., Fu Z., Jung S.Y., Jeong E.M, & Kim D.K. (2023). Purification and Characterization of Mitochondrial Mg2+-Independent Sphingomyelinase from Rat Brain. Molecules and Cells, 46(9), 545-557.

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Fisetin and d-Galactose Neuroprotection

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Fisetin (Lot#SLBF3913V) and d-galactose (sc-202564) were procured from Sigma-Aldrich Chemical Co. (St. Louis, MO, United States). The drugs were dissolved in 0.1% dimethyl sulfoxide (DMSO) and the final volume was adjusted with normal saline (0.9% saline). The control mice were injected with normal saline.
The antibodies used in the current studies are: SIRT1 (sc-74465), anti-Nrf2 (sc-722), anti-HO-1 (sc-136,961), P-JNK (sc-625), anti-p-NF-κβ (sc-136,548), anti-Iba-1 (sc-32,725),anti-GFAP (sc-33673) anti-interleukin (IL-1β) (sc-32,294), anti-tumor necrosis factor-α (TNF-α) (sc-52,746), NOS-2 (sc-651), anti-Bax (sc-7480) anti-Bcl2 (sc-7382), cleaved Caspase-3 (sc-7272), anti-PARP-1 (sc-8007), anti-PSD-95 (sc-71,933), anti-synaptosomal-associated protein 23 (SNAP-25), and anti-β-actin (sc-47,778) (Santa Cruz Biotechnology, Dallas, TX, United States). The primary antibodies were diluted in 1× TBST (1:1,000), and secondary anti-mouse HRP (horseradish peroxidase) conjugated (Promega Ref# W402) and anti-rabbit HRP conjugated (Promega Ref# W401) were diluted 1:10,000 in 1 M TBST (Promega, Fitchburg, WI, United States); the secondary antibodies (anti-mouse Ref# A11029 and anti-rabbit Ref# 32,732) used in the immunofluorescence studies were diluted in 1:100 in 1 M PBS.

Ahmad S., Khan A., Ali W., Jo M.H., Park J., Ikram M, & Kim M.O. (2021). Fisetin Rescues the Mice Brains Against D-Galactose-Induced Oxidative Stress, Neuroinflammation and Memory Impairment. Frontiers in Pharmacology, 12, 612078.

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Western Blot Analysis of Synaptic Proteins

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Synaptic fractions were prepared as above. Hippocampal lysates were prepared by homogenizing flash frozen tissue in RIPA buffer containing protease inhibitors. Following BCA assay (Thermo Fisher), 40–60μg of lysate or fraction was separated through 8% or 4–20% gradient polyacrylamide gels and transferred to nitrocellulose membranes (Thermo Fisher). Membranes were blocked with 5% non-fat dry milk in Tris buffered saline with 0.1% Tween 20 (TBST). Membranes were incubated in primary antibody (in 0.3% BSA in TBST) at 4°C overnight using antibodies to detect KL (KL2-34 (Maltare et al., 2014 (link))), β-tubulin (Developmental Studies Hybridoma Bank, E7, Iowa City, IA), synaptophysin (Abcam, AB52636, Cambridge, MA), GluN1 (Neuromab, 75–272, Davis, CA) and SNAP25 (SC73044, Santa Cruz). Secondary antibodies conjugated to HRP were detected by chemiluminescence (Immobilon, Millipore, Billerica, MA) and exposure to film.

Li Q., Vo H.T., Wang J., Fox-Quick S., Dobrunz L.E, & King G.D. (2017). Klotho regulates CA1 hippocampal synaptic plasticity. Neuroscience, 347, 123-133.

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Immunofluorescence Staining of HSV-1 Capsids

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Rabbit antibody against purified HSV-1 nuclear C capsids (PTNC) (1:5500 dilution) was kindy provided by Dr. Frazer Rixon, (MRC Virology Unit, Institute of Virology, UK) [19 (link)]. Mouse monoclonal anti-non-muscle myosin IIB (1:100 dilution, ab684) was obtained from Abcam, UK. Goat polyclonal anti-myosin V (1:200 dilution, LS-C139627) was obtained from Lifespan Bioscience, USA. Rabbit polyclonal anti-kinesin family member 3A (KIF3A, 1:50 dilution, SC-50457), rabbit polyclonal anti-Rab6 (1:50 dilution, SC-310) and mouse monoclonal anti-synaptosome associated protein 25 (SNAP25, 1:20 dilution, SC-20038) were obtained from Santa Cruz, USA. Alexa Fluor 633 Phalloidin and Alexa Fluor-labelled secondary antibodies were obtained from Thermo Fisher Scientific, USA.

Danastas K., Larsen A., Jobson S., Guo G., Cunningham A.L, & Miranda-Saksena M. (2022). Herpes simplex virus-1 utilizes the host actin cytoskeleton for its release from axonal growth cones. PLoS Pathogens, 18(1), e1010264.

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Molecular Markers of Synaptic Plasticity

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Antibodies directed against AMPKα1/α2 (#2603), ACC (#3676), phospho-ACC-Ser79 (#3661), PSD-95 (#2507), GluA1 (#13185), Raptor (#2280), phospho-Raptor-Ser792 (#2083), ULK1 (#8054), phospho-ULK1-Ser555 (#5869), SQSTM1/p62 (#5114), and LC3B (#2775) were obtained from Cell Signaling Technology. Anti-AMPKα2 (AF2850) was from R&D Systems. Anti-phospho-AMPK-Thr172 (sc-33524), synapsin Ia/IIb (sc-376622), Homer 1bc (sc-25271), synaptophysin (SYP) (sc-17750), GluN1 (NMDA zeta1; sc-1467), and SNAP25 (sc-376713) were from Santa Cruz Biotechnology. Anti-actin (#612656) and Munc (#610336) antibodies were from BD Transduction Laboratory. MAP2 (#M1406) antibody was from Sigma. SMI-312 (#837904) antibody was from Biolegend. AICAR and MRT68921 were purchased from Tocris, GSK621 was from Selleckchem, and Bafilomycin A1 was from Cell Signaling.

Domise M., Sauvé F., Didier S., Caillerez R., Bégard S., Carrier S., Colin M., Marinangeli C., Buée L, & Vingtdeux V. (2019). Neuronal AMP-activated protein kinase hyper-activation induces synaptic loss by an autophagy-mediated process. Cell Death & Disease, 10(3), 221.

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Antibody Profiling for Cell Signaling

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Anti–IL-1RA and VAMP5 antibodies were purchased from Abcam. Anti-Fas antibody was purchased from Millipore. Anti–Fap-1, Cav-1, SNAP25, CD105, CD90, CD44, CD63, CD9, and CD81 antibodies were purchased from Santa Cruz Biotechnology. Anti–p-NF-κB p65 and NF-κB p65 antibodies were purchased from Cell Signaling. Anti–β-actin antibody was purchased from Sigma-Aldrich. Alexa Fluor 488 and Alexa Fluor 568 secondary antibodies were purchased from Invitrogen. Protein A/G PLUS-Agarose was purchased from Santa Cruz Biotechnology. NF-κB inhibitor ammonium PDTC was purchased from Sigma-Aldrich.

Kou X., Xu X., Chen C., Sanmillan M.L., Cai T., Zhou Y., Giraudo C., Le A, & Shi S. (2018). The Fas/Fap-1/Cav-1 complex regulates IL-1RA secretion in mesenchymal stem cells to accelerate wound healing. Science translational medicine, 10(432), eaai8524.

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