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Synaptophysin

Synaptophysin is a membrane glycoprotein found in synaptic vesicles of neuronal and neuroendocrine cells.
It is widely used as a marker for synaptic density and neuroendocrine differentiation.
Synatophysin plays a key role in synaptic vesicle formation, exocytosis, and recycling.
Accurate quantification and localization of synaptophysin is crucial for understading synaptic function and neurodegenerative disorders.
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Most cited protocols related to «Synaptophysin»

The institutional review board of the University of Cologne approved this study. We collected and analysed fresh-frozen tumour samples of 152 SCLC patients, which were provided by multiple collaborating institutions as fresh-frozen tissue specimen, frozen sections or as genomic DNA extracted from fresh-frozen material (Extended Data Fig. 1). Human tumour samples were obtained from patients under IRB-approved protocols following written informed consent.
The fresh-frozen SCLC samples were primary tumours diagnosed as stage I–IV tumours, and snap-frozen after tissue sampling. All tumour samples were pathologically assessed to have a purity of at least 60% and no extensive signs of necrosis. Additionally, these tumour samples were reviewed by at least two independent expert pathologists and the diagnosis of SCLC was histomorphologically confirmed by H&E staining and immunohistochemistry for chromogranin A, synaptophysin, CD56 and Ki67. Matching normal material was provided in the form of EDTA-anticoagulated blood or adjacent non-tumorigenic lung tissue (Supplementary Table 1). The matched normal tissue was confirmed to be free of tumour contaminants by pathological assessment. Furthermore, tumour and matching normal material were confirmed to be acquired from the same patient by short tandem repeat (STR) analysis conducted at the Institute of Legal Medicine at the University of Cologne (Germany), or confirmed by subsequent SNP 6.0 array and sequencing analyses. Patient material was stored at −80 °C.
Whole-genome sequencing was performed on 110 SCLC fresh-frozen tumour samples and matched normal material. Additionally, we analysed RNA-seq data of 81 SCLC primary tumours (Extended Data Fig. 1 and Supplementary Table 1), among which 20 cases were previously published2 (link),43 . Furthermore, we studied the copy-number alterations of a total of 142 fresh-frozen tumour specimen by Affymetrix SNP 6.0, among which 74 cases were described before44 .
Clinical correlation studies were performed with the study cohort of 110 SCLC patients considering age of diagnosis, gender, tumour stage, surgery, treatment with chemotherapeutics, smoking status, smoking history and overall survival (Extended Data Figs 2 and 4 and Supplementary Table 1). The median follow-up time for this cohort of 110 SCLC patients was 69 months, and 31% of the patients were alive at the time of last follow-up (Extended Data Fig. 2a and Supplementary Table 1). Smoking status was available for 88% (n = 97) of the patients; 63% (n = 69) reported a smoking history amounting to a median of 45 pack-years. Patients with a known smoking history were further subcategorized to heavy smokers (>30 pack-years), average smokers (10–30 pack-years) and light/never smokers (<10 pack-years).
Primary findings on somatic mutations were further studied in a second independent cohort consisting of 112 SCLC cases. This validation cohort refers to the exome sequencing data of 28 fresh-frozen SCLC primary tumours and 9 SCLC cell lines2 (link),3 (link) which were re-analysed in this present study (Supplementary Table 7). Additionally, we performed targeted sequencing on 8 fresh-frozen and 67 formalin fixed paraffin embedded (FFPE) samples from SCLC patients (Supplementary Table 1).
Publication 2015
BLOOD Cells Chromogranin A Copy Number Polymorphism Diagnosis Diploid Cell Edetic Acid Formalin Freezing Frozen Sections Gender Genome Homo sapiens Immunohistochemistry Light Lung Mutation Necrosis Neoplasms Neoplastic Cell Transformation Operative Surgical Procedures Paraffin Embedding Pathologists Patients Pharmacotherapy RNA-Seq Short Tandem Repeat Small Cell Lung Carcinoma Synaptophysin Tissues

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Publication 2018
Adenocarcinoma BLOOD Carcinoma Carcinoma, Neuroendocrine Cells Chromogranins Chronic Obstructive Airway Disease Colonic Neoplasms Diploid Cell DNA Methylation Genome Germ Line Homo sapiens Males Malignant Neoplasms Mutation Patients Radiotherapy Synaptophysin Tissues X-Ray Photoelectron Spectroscopy
After three weeks withdrawal from chronic cocaine or saline treatment mice were decapitated. The ventral striatum was dissected from the brain as described elsewhere (Szumlinski et al., 2004 (link)), and subcellular fractionation was performed as described previously with minor modifications (Toda et al., 2006 (link)). Briefly, fresh brain tissues were homogenized in cold buffer containing 0.32 M sucrose and 10 mM HEPES, pH 7.4. Homogenates were cleared two times at 1000 g for 10 min to remove nuclei and large debris (P1). The resulting supernatants were concentrated at 12 000 g for 20 min to obtain a crude membrane fraction (P2), which was rinsed twice (4 mM HEPES, 1 mM EDTA, pH=7.4; 20 min at 12 000g). Then, it was incubated (20 mM HEPES, 100 mM NaCl, 0.5% triton X, pH= 7.2) for 15 min and centrifuged at 12 000 g for 20 min to pellet the synaptosomal membrane fraction (LP1). The supernatant was considered the non-postsynaptic density membrane fraction (non-PSD), sometimes referred to as the triton soluble fraction. The pellet was then solubilized (20 mM HEPES, 0.15 mM NaCl, 1% triton X100, 1% deoxycholic acid, 1% SDS, pH= 7.5) for 1 h and centrifuged at 10 000 for 15min. The supernatant contained the postsynaptic density fraction (PSD) or triton insoluble fraction. The integrity of non-PSD and PSD fractions was verified by immunoblotting for PSD-95 which was enriched in PSD fraction, and synaptophysin which was enriched in non-PSD fraction (Supplemental Figure 1). All buffers were supplemented with protease inhibitors cocktail (Complete mini tablets, Roche). Protein concentration was measured using the Bradford assay (Pierce).
Publication 2008
Biological Assay Brain Buffers Cell Nucleus Cocaine Cold Temperature Deoxycholic Acid Edetic Acid HEPES Mice, House Post-Synaptic Density Protease Inhibitors Proteins Radiotherapy Dose Fractionations Saline Solution Sodium Chloride Sucrose Synaptophysin Synaptosomes Tissue, Membrane Tissues Triton X-100 Ventral Striatum

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Publication 2014
Antibodies Argon Axon Cholinergic Receptors Cross Reactions Denervation Dental Plaque Dyes Fluorescence Forceps Helium Neon Gas Lasers IgG1 Immunoglobulins Laser Scanning Microscopy Light Mice, House Microscopy Muscle Tissue Nerve Endings Nerve Tissue Neurofilaments Neuromuscular Junction Submersion Synapses Synaptic Vesicles Synaptophysin tetramethylrhodamine Triton X-100 Z 300
For immunoblot analysis (14 ), the antibodies used were: p130 (BD Biosciences, 610261), p107 (Santa Cruz, SC-318), Synaptophysin (Neomarkers, RB-1461-P1), Karyopherin beta 1 (Santa Cruz, SC-1919), and α-Tubulin (Sigma, T9026).
Paraffin sections were rehydrated in Trilogy reagent (Cell Marque). Sections were mounted in ProLong® Gold Antifade reagent (Invitrogen) after immunostaining (14 ). The primary antibodies used were: Phospho-Histone 3 Ser10 (PH3) (Millipore, 06-570), BrdU (Becton-Dickinson, 347580), Ki67 (BD Biosciences, 55069), MCM6 (Santa Cruz Biotechnology, sc-9843), PCNA (Santa Cruz Biotechnology, sc-56), Surfactant protein C (SP-C) (Dr. Jeff Whitsett, University of Cincinnati), Clara cell secretory protein (CCSP) (Santa Cruz Biotechnology, sc-9772), Synaptophysin (SYP) (Neomarkers, RB-1461-P1). Alexa Fluor® secondary antibodies (Invitrogen) were used for antibody detection. Quantification was performed using the Bio-Quant image analysis software.
Publication 2010
alpha-Tubulin Antibodies Bromodeoxyuridine Cells Gold Histones Immunoblotting Immunoglobulins KPNB1 protein, human MCM6 protein, human Paraffin Proliferating Cell Nuclear Antigen Protein C Pulmonary Surfactants RBL1 protein, human Synaptophysin Uteroglobin

Most recents protocols related to «Synaptophysin»

Authorizations for reporting these three cases were granted by the Eastern Ontario Regional Forensic Unit and the Laboratoire de Sciences Judiciaires et de Médecine Légale du Québec.
The sampling followed a relatively standardized protocol for all TBI cases: samples were collected from the cortex and underlying white matter of the pre-frontal gyrus, superior and middle frontal gyri, temporal pole, parietal and occipital lobes, deep frontal white matter, hippocampus, anterior and posterior corpus callosum with the cingula, lenticular nucleus, thalamus with the posterior limb of the internal capsule, midbrain, pons, medulla, cerebellar cortex and dentate nucleus. In some cases, gross pathology (e.g. contusions) mandated further sampling along with the dura and spinal cord if available. The number of available sections for these three cases was 26 for case1, and 24 for cases 2 and 3.
For the detection of ballooned neurons, all HE or HPS sections, including contusions, were screened at 200×.
Representative sections were stained with either hematoxylin–eosin (HE) or hematoxylin-phloxin-saffron (HPS). The following histochemical stains were used: iron, Luxol-periodic acid Schiff (Luxol-PAS) and Bielschowsky. The following antibodies were used for immunohistochemistry: glial fibrillary acidic protein (GFAP) (Leica, PA0026,ready to use), CD-68 (Leica, PA0073, ready to use), neurofilament 200 (NF200) (Leica, PA371, ready to use), beta-amyloid precursor-protein (β-APP) (Chemicon/Millipore, MAB348, 1/5000), αB-crystallin (EMD Millipore, MABN2552 1/1000), ubiquitin (Vector, 1/400), β-amyloid (Dako/Agilent, 1/100), tau protein (Thermo/Fisher, MN1020 1/2500), synaptophysin (Dako/Agilent, ready to use), TAR DNA binding protein 43 (TDP-43) ((Protein Tech, 10,782-2AP, 1/50), fused in sarcoma binding protein (FUS) (Protein tech, 60,160–1-1 g, 1/100), and p62 (BD Transduc, 1/25). In our index cases, the following were used for the evaluation of TAI: β-APP, GFAP, CD68 and NF200; for the neurodegenerative changes: αB-crystallin, NF200, ubiquitin, tau protein, synaptophysin, TDP-43, FUS were used.
For the characterization of the ballooned neurons only, two cases of fronto-temporal lobar degeneration, FTLD-Tau, were used as controls. One was a female aged 72 who presented with speech difficulties followed by neurocognitive decline and eye movement abnormalities raising the possibility of Richardson’s disorder. The other was a male aged 67 who presented with a primary non-fluent aphasia progressing to fronto-temporal demαentia. In both cases, the morphological findings were characteristic of a corticobasal degeneration.
Publication 2023
Amyloid beta-Protein Precursor Amyloid Proteins Antibodies Broca Aphasia Cloning Vectors Congenital Abnormality Contusions Corpus Callosum Cortex, Cerebellar Cortex, Cerebral Corticobasal Degeneration Crystallins Dura Mater Eosin Eye Abnormalities Eye Movements Frontotemporal Lobar Degeneration FUBP1 protein, human Glial Fibrillary Acidic Protein Hematoxylin Immunohistochemistry Internal Capsule Iron Males Medial Frontal Gyrus Medulla Oblongata Mesencephalon Movement Movement Disorders neurofilament protein H Neurons Nucleus, Dentate Nucleus, Lenticular Occipital Lobe Periodic Acid phloxine Pons Proteins protein TDP-43, human RNA-Binding Protein FUS Saffron Sarcoma Seahorses Speech Spinal Cord Staining Synaptophysin Temporal Lobe Thalamus Ubiquitin White Matter Woman
The surgically removed brain tissue specimens were fixed with 3.7% neutral formaldehyde, embedded in conventional paraffin, sliced into 5-μm thick sections, and then stained with hematoxylin and eosin (H&E). Immunohistochemical staining was performed with the following primary antibodies: anti-BRAFV600E (Spring Bioscience, monoclonal, clone VE1, 1:50), glial fibrillary acidic protein (GFAP; Dako, 1:1000), neuronal nuclear antigen (NeuN; Chemicon, 1:2000), neurofilament (NF; Zymed, 1:100), synaptophysin (Biogenics, 1:50), oligodendrocyte transcription factor 2 (Olig-2; Immuno-Biological Laboratories, 1:500), epithelial cell membrane antigen (EMA; Zymed, 1:100), Ki-67 (MIB-1; OriGene, monoclonal, clone UMAB107, 1:200), p53 (Zymed, 1:100), CD68 (Bio-Rad, 1:500), CD34 (Zymed, monoclonal, clone QBEnd 10, 1:50), isocitrate dehydrogenase (IDH1/2; Zymed, 1:1000).
Histopathological records were systematically reviewed by two experienced neuropathologists according to the WHO classification scheme from 2016, including a panel of immunohistochemical markers. Ki-67 proliferation index was defined by the percentage of Ki-67 positive cells in the total cell population at 40 magnifications for 10 microscopic fields. In particular, tumors with IHC detection of CD34 expression and BRAFV600E mutation were analyzed in terms of their associations with clinical and pathological features.
Publication 2023
Antibodies Antigens, Nuclear Biopharmaceuticals Brain CA 15-3 Antigen Cells Clone Cells Eosin Formaldehyde Glial Fibrillary Acidic Protein Isocitrate Dehydrogenase (NAD+) Ki-67 Antigen Microscopy Mutation Neoplasms Neurofilaments Neurons Neuropathologist OLIG2 protein, human Operative Surgical Procedures Paraffin Embedding Synaptophysin Tissues
Western blot analysis was performed as described [21 (link)]. Tissues were harvested from control, S-W, and MSC groups (12 hours before surgical wound) at 6 hours after surgical wound. Anti-Claudin-5 antibody (Invitrogen, USA), anti-Occludin antibody (Abcam, USA), anti-ZO-1 antibody (Abcam, USA), anti-matrix metalloproteinase-9 (MMP-9) antibody (Abcam, USA), anti-synaptophysin antibody (Abcam, USA), and anti-endothelial nitric oxide (eNOS) antibody (Abcam, USA) were performed in the analysis. The ratio of the levels of targeted proteins to the level of β-actin was performed to analyze the trend of those proteins in the brain tissues.
Publication 2023
Actins anti-synaptophysin Antibodies, Anti-Idiotypic Brain Claudin-5 Endothelium Immunoglobulins Matrix Metalloproteinase 9 Occludin Oxide, Nitric Proteins Surgical Wound Synaptophysin Tissues Western Blot
ITSCs were seeded on roughened cover glasses with a density of 5 × 104 Cells. Immunocytochemistry was done as previously described in Ruiz-Perera et al. [33 (link)]. Afterwards, Primary antibodies against RELA (mouse, 200,301,065, 1:5000, Rockland, PA. USA), RELB (rabbit, D7D7W, 1:500, Cell Signaling Technology, Danvers, USA), c-Rel (rabbit, 4727, 1:500, Cell Signaling Technology), vGlut-II (rabbit, 07-1402-I, 1:300, Merck Millipore, Burlington, USA), NF200 (mouse, SAB3200747, 1:200, Sigma Aldrich), MAP2 (mouse, sC-390,543, 1:500, Santa Cruz, Dallas, USA) and synaptophysin (rabbit, ab32127, 1:500, Abcam, Cambridge, UK) were incubated for 1 h at RT. Secondary fluorochrome-conjugated antibodies (Alexa 488 anti-rabbit, Alexa 555 anti-mouse, Alexa 555 anti-rabbit, 1:300, Life Technologies, Germany) were incubated for 1 h at RT under the exclusion of light, followed by DAPI nuclear counterstaining. Fluorescence microscopy was performed using the confocal laser scanning microscope LSM 780 (Carl Zeiss, Jena, Germany). Randomly placed pictures were analyzed using ImageJ [34 ]. Nuclear fluorescence intensity was determined using the “measure” function in ImageJ on the nuclei of the cells.
Publication 2023
Antibodies Cell Nucleus DAPI Eyeglasses Fluorescence Fluorescent Dyes Immunocytochemistry Light METAP2 protein, human Microscopy, Confocal Microscopy, Fluorescence Mus Rabbits RELA protein, human rel Oncogene Synaptophysin
Mice were sacrificed and transcardially perfused with 4% paraformaldehyde (PFA) in cold phosphate-buffered saline (PBS). Mouse brains, superior cervical ganglions, and adrenal glands were collected, post-fixed in 4% PFA/PBS solution overnight, submerged in 30% sucrose in PBS for at least 72 h, and sectioned at 40 μm thickness using CM1950 cryostat (Leica)22 (link). Frozen sections were stained with antibodies specific to p150Glued (amino acid 3–202 at the N-terminus of p150Glued, BD Biosciences, #610474, 1:200, recognizing p150Glued but not p135+), p150Glued & p135+ (amino acid 1266–1278 at the C-terminus of p150Glued, Abcam, #ab11806, 1:500, recognizing both p150Glued and p135+), tyrosine hydroxylase (TH, Pel-Freez, #P40101-150, 1:2500; ImmunoStar, #22941, 1:500; Synaptic Systems, #213104, 1:500), dopamine transporter (DAT, Millipore, #MAB369, 1:500), vesicular monoamine transporter 2 (VMAT2, Synaptic Systems, #138302, 1:1000), glial fibrillary acidic protein (GFAP, Abcam, #ab7260, 1:1000), TAR DNA-binding protein 43 (TDP-43, Proteintech, #10782-2-AP, 1:500), α-synuclein (Santa Cruz, #sc-7011-R, 1:500; Santa Cruz, #sc-69977, 1:500), phosphorylated α-synuclein (Ser129) [p-α-synuclein (Ser129), Abcam, #ab51253, 1:500], neuronal nuclei (NeuN, Millipore, #ABN91, 1:500), synaptophysin (Millipore, #AB9272, 1:500), binding immunoglobulin protein (BiP, also referred to as GRP78, Abcam, #ab21685, 1:500), reticulon 3 (RTN3, Proteintech, #12055-2-AP, 1:500), 63 kDa cytoskeleton-linking membrane protein (CLIMP63, Proteintech, #16686-1-AP, 1:500), calnexin (Abcam, #ab22595, 1:500), protein disulfide isomerase (PDI, Proteintech, #11245-1-AP, 1:500), receptor binding cancer antigen expressed on SiSo cells (RCAS1, Cell Signaling Technology, #12290, 1:500), early endosome antigen 1 (EEA1, Cell Signaling Technology, #3288, 1:500), sequestosome 1 (SQSTM1, MBL, #PM066, 1:500), cathepsin D (R&D Systems, #AF1029, 1:500), ER-Golgi intermediate compartment 53 kDa protein (ERGIC53, Sigma-Aldrich, #E1031, 1:500), 130 kDa cis-Golgi matrix protein (GM130, BD Biosciences, #610822, 1:500), phosphorylated eukaryotic translation initiation factor 2α (Ser51) [p-eIF2α (Ser51), Abcam, #ab32157, 1:500], and phosphorylated inositol-requiring enzyme 1α (Ser724) [p-IRE1α (Ser724), Abcam, #ab48187, 1:500] as suggested by manufacturers. Alexa Fluor 488-, 546-, or 647-conjugated secondary antibody (Invitrogen, 1:500) was used to visualize the staining. Fluorescent images were captured using LSM 880 laser-scanning confocal microscope with Zen software (Zeiss) in conventional or Airyscan mode. As a high-resolution imaging modality, the Airyscan technology is reported to improve resolution 2-fold and signal-to-noise ratio 8-fold relative to the conventional confocal microscopy61 (link). The paired images in all the figures were collected at the same gain and offset settings. Post-collection processing was applied uniformly to all paired images. The images were presented as a single optic layer after acquisition in z-series stack scans at 1.0 μm intervals from individual fields or displayed as maximum-intensity projection or three-dimensional (3D) reconstruction to represent confocal stacks.
Publication 2023
Adrenal Glands alexa fluor 488 alpha-Synuclein Amino Acids Antibodies Antigens Binding Proteins Brain Calnexin Cathepsin D Cell Nucleus Cells Cold Temperature Cytoskeleton Dopamine Transporter dynactin subunit 1, human early endosome antigen 1 Enzymes ERN1 protein, human Eukaryotic Initiation Factors Eye Frozen Sections Ganglia, Superior Cervical Glial Fibrillary Acidic Protein Glucose Regulated Protein 78 kDa Golgi Apparatus Golgin subfamily A member 2 Immunoglobulins Inositol Malignant Neoplasms Membrane Proteins Microscopy, Confocal Mus Neurons paraform Phosphates Protein Disulfide-Isomerases Proteins protein TDP-43, human Radionuclide Imaging Reconstructive Surgical Procedures Saline Solution SQSTM1 protein, human Sucrose Synaptophysin Tyrosine 3-Monooxygenase Vesicular Monoamine Transporter 2

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Synaptophysin is a protein marker used in immunohistochemical analysis to detect synaptic vesicles in neuronal tissues. It is a widely used tool for the identification and characterization of neuronal and neuroendocrine cells.
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Synaptophysin is a membrane glycoprotein found in presynaptic vesicles of neuronal and neuroendocrine cells. It is a commonly used marker for the identification of neuroendocrine and neuronal cells.
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PSD95 is a protein that functions as a scaffold in the postsynaptic density of neuronal synapses. It plays a role in the organization and clustering of neurotransmitter receptors, ion channels, and signaling molecules at the postsynaptic membrane.
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NeuN is a protein marker used for the detection and identification of neuronal cell nuclei in various vertebrate species. It is commonly used in immunohistochemistry and other laboratory techniques to study the distribution and properties of neurons in biological samples.
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PSD95 is a protein that functions as a scaffold, organizing and anchoring various signaling molecules at the postsynaptic density of neuronal synapses. It plays a crucial role in the organization and regulation of synaptic structure and function.
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β-actin is a protein that is found in all eukaryotic cells and is involved in the structure and function of the cytoskeleton. It is a key component of the actin filaments that make up the cytoskeleton and plays a critical role in cell motility, cell division, and other cellular processes.
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Synaptophysin is a glycoprotein found in the membrane of synaptic vesicles. It is a commonly used biomarker for the identification and quantification of synapses in both neurological and non-neurological tissues.
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Synaptophysin is a protein found in the synaptic vesicles of presynaptic neurons. It is commonly used as a marker in immunohistochemistry and Western blotting applications to identify the presence and distribution of synaptic terminals.
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More about "Synaptophysin"

synapse, neurodegenerative, neuroendocrine, PSD95, NeuN, β-actin, Alexa Fluor 488, Ab32127, synaptic vesicle, exocytosis, reproducibility, accuracy