Vglut

Manufactured by Synaptic Systems

VGLUT is a protein that functions as a vesicular glutamate transporter. It is responsible for the uptake of glutamate into synaptic vesicles, which is a crucial step in the release of this neurotransmitter at presynaptic terminals.

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Lab products found in correlation

4 protocols using vglut

Protein Quantification and Western Blot

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Protein samples were lysed in RIPA lysis buffer (Beyotime Institute of Biotechnology) with protease inhibitor cocktail and the protein concentration was measured using the BCA kit (Shanghai Yeasen Biotechnology). The Western blot was performed as previously described using vGLUT (1:1000, Synaptic System) and GAPDH (1:4000, Kangchen) as primary antibodies.

Cao S.Y., Hu Y., Chen C., Yuan F., Xu M., Li Q., Fang K.H., Chen Y, & Liu Y. (2017). Enhanced derivation of human pluripotent stem cell-derived cortical glutamatergic neurons by a small molecule. Scientific Reports, 7, 3282.

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Immunohistochemical Analysis of Neuroendocrine Markers

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Primary antibodies used are: rabbit anti-GnRH (Beauvillain and Tramu, 1980 (link)), sheep anti-KP AC053 (Franceschini et al., 2013 (link)), sheep anti-GnRH (Skrapits et al., 2015 (link)), mouse anti-p140Cap (Grasso et al., 2018 (link), see Supplementary Figure 1 for specificity test), guinea pig anti-Vesicular GABA Transporter (VGAT, Synaptic System, used 1:500), guinea pig anti-Vesicular glutamate Transporter (VGLUT, Synaptic System, used 1:500), rabbit anti-c-Fos (sc-52 Santacruz, used 1:500) and rabbit anti-cleaved caspase 3 (Cell Signaling Technology, used 1:400, Whittaker et al., 2021 (link)). Secondary antibodies used are: Alexa-Fluor 488 goat anti-rabbit (Invitrogen, used 1:400), Cy3 goat anti-rabbit (Invitrogen, used 1:800), Alexa-Fluor 568 goat anti-mouse (Invitrogen, used 1:400), FITC anti-guinea pig (Sigma F6261, used 1:100), Cy3 donkey anti-sheep (Jackson, used 1:1000). Peroxidase-conjugated secondary antibodies were obtained from GE Healthcare.

Camera M., Russo I., Zamboni V., Ammoni A., Rando S., Morellato A., Cimino I., Angelini C., Giacobini P., Oleari R., Amoruso F., Cariboni A., Franceschini I., Turco E., Defilippi P, & Merlo G.R. (2022). p140Cap Controls Female Fertility in Mice Acting via Glutamatergic Afference on Hypothalamic Gonadotropin-Releasing Hormone Neurons. Frontiers in Neuroscience, 16, 744693.

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Molecular Markers of Synaptic Signaling

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PI3K-C2β (BD Biosciences, #611342, 1:1000), phospho-p70 S6 kinase (Thr389; Cell Signaling, #9205, 1:500), p70 S6 kinase (Cell Signaling, #9202, 1:1000), S6 ribosomal protein (Cell Signaling, #2217, 1:1000), phospho-S6 ribosomal protein (Ser235/236; Cell Signaling, #2211, 1/1000), phospho-Akt (Ser473) antibody (Cell Signaling, #9271, 1:1000), Akt (pan; Cell Signaling, #4691, 1:1000), GAPDH (Cell Signaling, #2118, 1/2500), PSD95 (Synaptic Systems, #124 011, 1:1000), vGAT (Synaptic Systems, #131011, 1:1000), vGLUT (Synaptic Systems, #135302, 1:1000), synaptotagmin 1 (1Synaptic Systems, #105 011, 1:5000), mTOR (Cell Signaling, #7C10, 1:1000), GluA1 (Millipore, # MAB2263, 1:1000), Homer1 (Synaptic Systems, #160003, 1:1000), Rab7 (Cell Signaling, #9367, 1:1000), p62 (Cell Signaling, #5114, 1:1000), LC3 (Abcam, #ab48394, 1:1000), GFP (Abcam, #ab290, 1:1000), Vinculin (Abcam, #ab129002, 1:5000).

Gozzelino L., Kochlamazashvili G., Baldassari S., Mackintosh A.I., Licchetta L., Iovino E., Liu Y.C., Bennett C.A., Bennett M.F., Damiano J.A., Zsurka G., Marconi C., Giangregorio T., Magini P., Kuijpers M., Maritzen T., Norata G.D., Baulac S., Canafoglia L., Seri M., Tinuper P., Scheffer I.E., Bahlo M., Berkovic S.F., Hildebrand M.S., Kunz W.S., Giordano L., Bisulli F., Martini M., Haucke V., Hirsch E, & Pippucci T. (2022). Defective lipid signalling caused by mutations in PIK3C2B underlies focal epilepsy. Brain, 145(7), 2313-2331.

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Neuronal Protein Localization Assay

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Gephyrin (1:200, Synaptic Systems, #147318), GFAP (1:500, Sigma, G3893), MAP2 (1:1000, Sigma-Aldrich, #M9942), phospho-S6 ribosomal protein (Ser235/236; 1:200, Cell Signaling, #2211), p62 (Progen, #GP62-C, 1:400), PSD95 (1:400, Synaptic Systems, #124 011), neurofilament M (1:200, Chemicon #AB1987), vGAT (1:400, Synaptic Systems, #131011C2), vGLUT (1:400, Synaptic Systems, #135302).

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