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Synaptotagmin

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Synaptotagmin is a membrane-bound protein that plays a central role in the regulation of synaptic vesicle exocytosis and neurotransmitter release. It senses changes in calcium concentration and triggers the fusion of synaptic vesicles with the presynaptic membrane, allowing the release of neurotransmitters into the synaptic cleft.

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Lab products found in correlation

β actin, by Abcam (2 mentions) Synaptophysin, by Santa Cruz Biotechnology (1 mentions) Ab8227, by Abcam (1 mentions) Spinophilin, by Abcam (1 mentions) Biomax film, by Kodak (1 mentions) Bca protein assay, by Avantor (1 mentions) Synapsin 1, by Abcam (1 mentions) Darpp32, by Abcam (1 mentions) Gabaarβ3, by Abcam (1 mentions) Psd95, by Abcam (1 mentions) Ecl reagent, by Thermo Fisher Scientific (1 mentions) Image studio lite, by LI COR (1 mentions) Odyssey near infrared fluorescence imaging system, by LI COR (1 mentions)

3 protocols using synaptotagmin

1

Quantifying Neuronal and Glial Markers

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Western blot was performed following determination of sample protein concentration using a BCA assay kit (Pierce, Thermo; Rockford, IL, USA). Membranes were probed with antibodies against glial fibrillary acidic protein (DAKO/Agilent Technologies, 1:1000), synaptophysin (Santa Cruz, #sc‐17750, 1:1000), synaptotagmin (Abcam, #ab51164 1:1000), PSD‐95 (CST, #3450, 1:1000), NR2B (Millipore, #06‐600, 1:1000) and β‐actin (Abcam, #ab8227, 1:5000).
Pang C.C., Sørensen M.H., Lee K., Luk K.C., Trojanowski J.Q., Lee V.M., Noble W, & Chang R.C. (2022). Investigating key factors underlying neurodegeneration linked to alpha‐synuclein spread. Neuropathology and Applied Neurobiology, 48(6), e12829.
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2

Western Blot Analysis of Neuronal Proteins

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The total amount of protein in the lysates was determined by BCA protein assay (AMRESCO). Samples (50 μg protein) were separated by 10% SDS-PAGE and electroblotted to PVDF membranes, which were blocked by incubation in 5% non-fat dry milk dissolved in TBS-T (150 mM NaCl, 50 mM Tris, 0.05% Tween 20). Following transfer, proteins were probed using the following primary antibodies: D1R, DARPP32, GABAARβ1, GABAARβ3, TrkB, pAKT, AKT, synaptotagmin, synapsin I, PSD95, spinophilin, and β-actin (Abcam). Then, horseradish peroxidase-conjugated secondary antibody was used. After extensive washing, protein bands detected by antibodies were visualized by ECL reagent (Thermo) after exposure on Kodak BioMax film (Kodak).
Ding S., Zhuge W., Hu J., Yang J., Wang X., Wen F., Wang C, & Zhuge Q. (2018). Baicalin reverses the impairment of synaptogenesis induced by dopamine burden via the stimulation of GABAAR–TrkB interaction in minimal hepatic encephalopathy. Psychopharmacology, 235(4), 1163-1178.
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3

Dot Blotting for CRPS Autoantigen Screening

Cited 1 time
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Recombinant human proteins were purchased for dot blotting to measure specific IgM binding reactivity in the sera of normal controls, early CRPS patients, chronic CRPS patients, and resolved CRPS patients. The 9 protein candidates initially screened included; 1) alpha enolase, 2) gamma actin, 3) glutamate ionotropic receptor delta type subunit 2 (GRID2), 4) eukaryotic translation elongation factor 1 alpha 1(EEF1A1), 5) beta tubulin, 6) interleukin 1 receptor associated kinase 1(IRAK1), and 7) synaptotagmin were all obtained from Abcam, Burlingame, CA, 8) keratin 16 (Novus Biologicals, Centennial, CO), and 9) histone 3.2 (New England Biolabs, Ipswich, MA). Full length proteins were utilized whenever commercially available. These potential autoantigens were selected based on results of our previous liquid chromatography-tandem mass spectroscopy studies in fracture mouse skin.[42 (link)] Each recombinant protein (2ul) was applied to a nitrocellulose membrane and incubated for 1 hour with human sera (1:1000 dilution in TPBS) followed by 1hour incubation with anti-human IgM-Dylight800 secondary antibody (1:10000 dilution, Thermo Fisher Scientific, Waltham, MA). The signals were detected using Odyssey Near-Infrared Fluorescence Imaging System and quantified using Image Studio Lite (LI-COR Biosciences, Lincoln, NE).
Guo T.Z., Wei T., Tajerian M., Clark J.D., Birklein F., Goebel A., Li W.W., Sahbaie P., Escolano F.L., Herrnberger M., Kramer H.H, & Kingery W.S. (2020). Complex regional pain syndrome patient IgM has pronociceptive effects in the skin and spinal cord of tibia fracture mice. Pain, 161(4), 797-809.
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