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Anti synaptophysin

Manufactured by Santa Cruz Biotechnology
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Anti-synaptophysin is a primary antibody used for the detection and visualization of synaptophysin, a membrane glycoprotein found in presynaptic vesicles. It is commonly used in immunohistochemical and immunocytochemical applications to identify and localize synaptic structures.

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Lab products found in correlation

Fluoro gel mounting medium, by Electron Microscopy Sciences (2 mentions) Mouse anti βiii tubulin, by Merck Group (2 mentions) Rabbit anti tom20, by Santa Cruz Biotechnology (2 mentions) Anti myo6, by Merck Group (2 mentions) Anti map2, by BD (2 mentions) Anti myo6, by Santa Cruz Biotechnology (2 mentions) Anti cyto c, by BD (2 mentions) Anti map2, by Cell Signaling Technology (1 mentions) Axio vert a1 microscope, by Zeiss (1 mentions) Alexa fluor 488 antibody, by Thermo Fisher Scientific (1 mentions) Anti apoa 1, by Santa Cruz Biotechnology (1 mentions) Anti synapsin 1, by Abcam (1 mentions) Anti syntaxin 1b, by Synaptic Systems (1 mentions) Anti apoe, by Abcam (1 mentions) Protein g sepharose 4 fast flow beads, by GE Healthcare (1 mentions) Vibratome, by Leica (1 mentions) Avidin biotin complex abc, by Vector Laboratories (1 mentions) Anti gfap, by Thermo Fisher Scientific (1 mentions) Anti aqp4, by Santa Cruz Biotechnology (1 mentions) Anti occludin, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Goat anti-synaptophysin Rabbit Anti-Synaptophysin Mouse anti-synaptophysin Synaptophysin

6 protocols using anti synaptophysin

1

Visualizing Hippocampal Neuron Synapses

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Primary hippocampal neurons fixed in 10% buffered formalin were blocked in 10% goat serum for 1 h, then incubated with anti-Bassoon (1:10, Santa Cruz Biotechnology, Dallas, TX, USA), anti-synaptophysin (1:10, Santa Cruz Biotechnology, Dallas, TX, USA), anti-MAP2 (1:100, Cell Signaling Technology, Danvers, MA, USA) antibodies overnight at 4 °C. Cells were washed and incubated with Alexa-fluor 488 antibody or Alexa-568 antibody (1:200 dilution; Invitrogen, Molecular Probes, Carlsbad, CA, USA) for 1 h at room temperature and mounted on glass slides. Images were taken with a Zeiss Axiovert A1 microscope and processed using AxioVision 4.9.
Jansen J., Scott M., Amjad E., Stumpf A., Lackey K.H., Caldwell K.A, & Park H.A. (2021). Bcl-xL Is Required by Primary Hippocampal Neurons during Development to Support Local Energy Metabolism at Neurites. Biology, 10(8), 772.
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2

Immunostaining of Cortical Neurons

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Cortical neurons were fixed with 4% paraformaldehyde and 4% sucrose at RT for 20 min, washed three times with PBS for 5 min each, permeabilized in 0.15% Triton X-100 for 15 min, and then incubated in blocking buffer (4% BSA and 3% normal goat serum) in PBS for 1 hr. Fixed neurons were next incubated with primary antibodies diluted in blocking buffer at 4°C overnight. Primary antibodies were as follows: rabbit anti-TOM20 (1:100, Santa Cruz), anti-myo6 (1:100, Sigma-Aldrich), anti-synaptophysin (1:400, Santa Cruz), and anti-SNPH (1:250); mouse anti-βIII-tubulin (1:5000, Sigma-Aldrich), anti-MAP2 (1:5000, BD Biosciences), anti-SV2 (1:2000, DSHB), anti-myo6 (1:50, Santa Cruz), and anti-cyto c (1:100, BD Biosciences). After washing three times with PBS at RT for 10 min each, samples were incubated with secondary fluorescent antibodies (Alexa 488, 546, 594 or 633 conjugated, Thermo Fisher Scientific) diluted in blocking buffer for 60 min, rewashed with PBS, and mounted with Fluoro-Gel mounting medium (Electron Microscopy Sciences) before imaging.
Li S., Xiong G.J., Huang N, & Sheng Z.H. (2020). The crosstalk of energy sensing and mitochondrial anchoring sustains synaptic efficacy by maintaining presynaptic metabolism. Nature metabolism, 2(10), 1077-1095.
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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.
Liu C., Song X., Nisbet R, & Götz J. (2016). Co-immunoprecipitation with Tau Isoform-specific Antibodies Reveals Distinct Protein Interactions and Highlights a Putative Role for 2N Tau in Disease. The Journal of Biological Chemistry, 291(15), 8173-8188.
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4

Immunostaining of Cortical Neurons

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Cortical neurons were fixed with 4% paraformaldehyde and 4% sucrose at RT for 20 min, washed three times with PBS for 5 min each, permeabilized in 0.15% Triton X-100 for 15 min, and then incubated in blocking buffer (4% BSA and 3% normal goat serum) in PBS for 1 hr. Fixed neurons were next incubated with primary antibodies diluted in blocking buffer at 4°C overnight. Primary antibodies were as follows: rabbit anti-TOM20 (1:100, Santa Cruz), anti-myo6 (1:100, Sigma-Aldrich), anti-synaptophysin (1:400, Santa Cruz), and anti-SNPH (1:250); mouse anti-βIII-tubulin (1:5000, Sigma-Aldrich), anti-MAP2 (1:5000, BD Biosciences), anti-SV2 (1:2000, DSHB), anti-myo6 (1:50, Santa Cruz), and anti-cyto c (1:100, BD Biosciences). After washing three times with PBS at RT for 10 min each, samples were incubated with secondary fluorescent antibodies (Alexa 488, 546, 594 or 633 conjugated, Thermo Fisher Scientific) diluted in blocking buffer for 60 min, rewashed with PBS, and mounted with Fluoro-Gel mounting medium (Electron Microscopy Sciences) before imaging.
Li S., Xiong G.J., Huang N, & Sheng Z.H. (2020). The crosstalk of energy sensing and mitochondrial anchoring sustains synaptic efficacy by maintaining presynaptic metabolism. Nature metabolism, 2(10), 1077-1095.
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5

Multimodal Labeling of Brain Sections

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Fixed brain hemispheres were cut into 40 µm sagittal sections with a vibratome (Leica). Sections were incubated with primary antibodies and were incubated overnight at 4°C, including anti-Aβ MOAB-2 (6C3) (Millipore) at 1:1000; and anti-GFAP (Invitrogen) at 1:500, anti-Iba1 (Wako) at 1:500; anti-synaptophysin (Santa Cruz) at 1:200, anti-pan-laminin (Sigma) at 1:300; anti-fibrinogen (Dako) at 1:2000; anti-AQP4 (Santa Cruz) at 1:100 or 1:200; anti-occludin (Invitrogen) at 1:100 in 2% normal horse serum in Tris-buffered saline (TBS)-Triton 0.02%. The following day, sections were incubated with appropriate secondary antibodies (1:400; Vector), followed by enhancement with avidin-biotin complex (ABC; Vector) and 3,3′-diaminobenzidine (DAB; Sigma). Images were acquired with a Leica DM2500 microscope connected to a camera (Q-Imaging Micropublisher 3.3 RTV) using Q-capture Pro software. For immunofluorescence, fluorescent secondary antibodies (1:400 Alexa Fluor®, Invitrogen) were used. In some cases, staining with 647-labelled tomato lectin (Vector Labs) was carried out simultaneously with secondary antibody incubation. For Thioflavin S staining, sections were incubated with 1% Thioflavin S (Sigma) for 8 min before being washed with ethanol and H2O.
Ries M., Watts H., Mota B.C., Lopez M.Y., Donat C.K., Baxan N., Pickering J.A., Chau T.W., Semmler A., Gurung B., Aleksynas R., Abelleira-Hervas L., Iqbal S.J., Romero-Molina C., Hernandez-Mir G., d’Amati A., Reutelingsperger C., Goldfinger M.H., Gentleman S.M., Van Leuven F., Solito E, & Sastre M. (2021). Annexin A1 restores cerebrovascular integrity concomitant with reduced amyloid-β and tau pathology. Brain, 144(5), 1526-1541.
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6

Rat Brain Synapse Protein Analysis

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Fifty male Wistar rats (280–300 g) were obtained from shanghai laboratory animal center, Chinese Academy of Sciences. Four rats were kept in a cage and allowed free access to water and food. The house was maintained at 22–25°C with a 12-hr light-dark cycle.
Mouse monoclonal anti-PSD-95 (postsynaptic density protein-95) and anti-synaptophysin were respectively obtained from Santa Cruz Biotechnology (sc-32290) and Abcam Inc (ab8049). Rabbit polyclonal anti-IBA (ionized calcium binding adapter) was purchased from Wako (019-19741). Mouse monoclonal anti-GFAP (Glial fibrillary acidic protein) was purchased from EMD Millipore (MAB3402). Anti-mouse IgG, HRP conjugate was purchased from Kang Chen Biotechnology (KC-MM-035). Anti-mouse IgG-FITC and anti-rabbit IgG-Rhodamine were purchased from Jackson Biotechnology (115-095-003, 111-025-003).
Tian X.S., Guo X.J., Ruan Z., Lei Y., Chen Y.T, & Zhang H.Y. (2014). Long-Term Vision and Non-Vision Dominant Behavioral Deficits in the 2-VO Rats Are Accompanied by Time and Regional Glial Activation in the White Matter. PLoS ONE, 9(6), e101120.
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