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Guinea pig anti vglut2

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Guinea pig anti-VGluT2 is a primary antibody used for the identification and localization of the VGluT2 (vesicular glutamate transporter 2) protein in biological samples. VGluT2 is a marker for glutamatergic neurons. This antibody can be used in various immunohistochemical and immunocytochemical applications.

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Lab products found in correlation

Rabbit anti gfp, by Thermo Fisher Scientific (3 mentions) Guinea pig anti vglut1, by Merck Group (3 mentions) Rabbit anti ha, by Cell Signaling Technology (2 mentions) Goat anti glud2, by Santa Cruz Biotechnology (2 mentions) Rabbit anti dsred, by Takara Bio (2 mentions) Mouse anti neun, by Merck Group (2 mentions) Goat anti chat, by Merck Group (2 mentions) Mouse anti cre, by Merck Group (2 mentions) Sheep anti gfp, by Bio-Rad (2 mentions) Goat anti wga, by Vector Laboratories (2 mentions) Mouse anti sv2, by Developmental Studies Hybridoma Bank (2 mentions) Mouse anti ctb, by Abcam (2 mentions) Living colors dsred, by Santa Cruz Biotechnology (2 mentions) Goat anti dtr, by Santa Cruz Biotechnology (2 mentions) Goat anti ctb, by List Biological Laboratories (2 mentions) Alexa fluor 647 goat anti guinea pig, by Jackson ImmunoResearch (2 mentions) Alexa 488 conjugated donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Mouse anti gfp, by Thermo Fisher Scientific (1 mentions) Confocal microscope, by Zeiss (1 mentions) Alexa 488 conjugated donkey anti mouse igg, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

VGluT2 (15 citations) Anti-VGluT2 (5 citations)

17 protocols using guinea pig anti vglut2

1

Immunohistochemistry for Neurotransmitter Markers

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Selected sections at the level of the dDG were processed using the MOM kit as described above. Sections were incubated overnight at RT in a solution containing rabbit anti-GFP (1:2000; Invitrogen), guinea pig anti-VGLUT2 (1:5000; Millipore) and mouse anti-GAD65 (1:100; Millipore) or mouse anti-GFP (1:100; Invitrogen), guinea pig anti-VGLUT2 (1:5000; Millipore) and rabbit anti-VGAT (1:1000; Synaptic System) diluted in MOM diluent. After several rinses in KPBS, they were incubated for 2 h in Alexa488-conjugated donkey anti-rabbit IgG (1:200; Invitrogen), Cy5-conjugated donkey anti-guinea pig (1:100; Jackson ImmunoResearch Laboratories, Inc.), and Cy3-conjugated donkey anti-mouse (1:100; Jackson ImmunoResearch Laboratories, Inc.) or Alexa488-conjugated donkey anti-mouse IgG (1:200; Invitrogen), Cy5-conjugated donkey anti-guinea pig (1:100; Jackson ImmunoResearch Laboratories, Inc.), and Cy3-conjugated donkey anti-rabbit (1:100; Jackson ImmunoResearch Laboratories, Inc.) diluted in MOM diluent. After several rinses in KPBS, all sections were then mounted on superfrost-coated slides, dried overnight at RT and coverslipped with Fluoromount. The specimens were analyzed with confocal microscope (Zeiss).
Billwiller F., Castillo L., Elseedy H., Ivanov A.I., Scapula J., Ghestem A., Carponcy J., Libourel P.A., Bras H., Abdelmeguid N.E., Krook-Magnuson E., Soltesz I., Bernard C., Luppi P.H, & Esclapez M. (2020). GABA–glutamate supramammillary neurons control theta and gamma oscillations in the dentate gyrus during paradoxical (REM) sleep. Brain Structure & Function, 225(9), 2643-2668.
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2

Immunostaining of Glutamate Receptors

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We used the following primary antibodies: goat anti-GluD2 (Santa Cruz Biotechnology or Frontier Institute; 1:200), rabbit anti-HA (Cell Signaling Technology; 1:500), guinea pig anti-vGluT1 (Millipore Sigma or Frontier Institute, 1:200), guinea pig anti-vGluT2 (Millipore Sigma, or Frontier Institute, 1:200). In some MADM experiments, chicken anti-GFP (Aves; 1:500), rabbit anti-DsRed (Clontech; 1:500) were used.
Takeo Y.H., Shuster S.A., Jiang L., Hu M., Luginbuhl D.J., Rülicke T., Contreras X., Hippenmeyer S., Wagner M.J., Ganguli S, & Luo L. (2020). GluD2- and Cbln1-mediated Competitive Interactions Shape the Dendritic Arbors of Cerebellar Purkinje Cells. Neuron, 109(4), 629-644.e8.
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3

Immunostaining of Glutamate Receptors

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We used the following primary antibodies: goat anti-GluD2 (Santa Cruz Biotechnology or Frontier Institute; 1:200), rabbit anti-HA (Cell Signaling Technology; 1:500), guinea pig anti-vGluT1 (Millipore Sigma or Frontier Institute, 1:200), guinea pig anti-vGluT2 (Millipore Sigma, or Frontier Institute, 1:200). In some MADM experiments, chicken anti-GFP (Aves; 1:500), rabbit anti-DsRed (Clontech; 1:500) were used.
Takeo Y.H., Shuster S.A., Jiang L., Hu M., Luginbuhl D.J., Rülicke T., Contreras X., Hippenmeyer S., Wagner M.J., Ganguli S., & Luo L. (2020). GluD2- and Cbln1-mediated Competitive Synaptogenesis Shapes the Dendritic Arbors of Cerebellar Purkinje Cells.
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4

Immunostaining and 3DISCO Protocols

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An overview of the antibodies used is presented in Table 1 and Table 2. For DAB staining, we used a primary rabbit anti-RFP antibody (1:1000, Rockland Immunochemicals, Limerick, PA, USA) and a secondary donkey anti-rabbit antibody (1:200, Jackson, Cambridge, UK). For immunofluorescence, we used chicken anti-calbindin (1:500, Synaptic Systems, Goettingen, Germany), mouse anti-calbindinD-28 (1:500, Sigma-Aldrich), chick anti-GFP (1:200, Abcam, Cambridge, UK), goat anti-FoxP2 (1:500, Santa Cruz, Santa Cruz, CA, USA), guinea pig anti-vGluT2 (1:500, MilliPore, Amsterdam, The Netherlands), rabbit anti-RFP (1:1000, Rockland) and mouse anti-NeuN (1:1000, MilliPore) as primary antibodies. Cy5 anti-chicken (1:200, Jackson), Cy3 anti-mouse, FITC anti-chick, (1:200, MilliPore), Alexa488 anti-goat (1:200, Jackson), Cy5 anti-guinea pig (1:200, Jackson), Cy3 anti-rabbit (1:400, Jackson) and Alexa488 anti-Mouse (1:200, Jackson) antibodies were used as secondary antibodies.
For 3DISCO, we used chicken anti-calbindin (1:500, Synaptic Systems), goat anti-FoxP2 (1:500, Santa Cruz) and rabbit anti-RFP (1:000, Rockland) as primary antibodies. Cy5 anti-chicken (1:200, Jackson), Alexa488 anti-goat (1:200, Jackson) and Cy3 anti-rabbit (1:400, Jackson) antibodies were used as secondary antibodies.
Dumas D.B., Gornati S.V., Adolfs Y., Shimogori T., Pasterkamp R.J, & Hoebeek F.E. (2022). Anatomical Development of the Cerebellothalamic Tract in Embryonic Mice. Cells, 11(23), 3800.
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5

Immunohistochemical Profiling of Neuronal Markers

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Immunohistochemistry was carried out using the following antibodies: mouse anti-Calbindin (CB, 1:3000, Sigma), rabbit anti-Calretinin (CR, 1:400, Invitrogen), mouse anti-Reelin (Reln, G10, 1:2000, provided by AMG), rabbit anti-Pax6 (1:200, Covance), mouse anti-neurofilament (NF; 2H3,1:500, Hybridoma bank), mouse anti-Isl1 (39.4D5, 1:500, Hybridoma bank), rabbit anti-Isl1 (1:2000, Abcam), goat-anti cleaved Caspase3 (1:500, bcam), goat-anti-choline acetyltransferase (ChAT, 1:100, Millipore) and rat-anti Fzd3 (1:400, R&D). Cortical barrels were studied in vibratome sections of flattened cortex, stained with guinea pig anti-Vglut2 (1:2000, Millipore). Signal was detected with an anti-mouse/rabbit universal ABC kit (PK-6200, Universal, Vector) or the following fluorescent secondary antibodies: donkey anti-mouse Alexa fluor 546 (1:1000, Invitrogen), donkey anti-mouse Alexa fluor 488 (1:1000, Invitrogen), donkey anti-rabbit Alexa fluor 488 (1:1000, Invitrogen), donkey anti-rabbit Alexa fluor 546 (1:1000, Invitrogen), goat anti-guinea pig Alexa fluor488(1:1000, Invitrogen).
Feng J., Xian Q., Guan T., Hu J., Wang M., Huang Y., So K.F., Evans S.M., Chai G., Goffinet A.M., Qu Y, & Zhou L. (2016). Celsr3 and Fzd3 Organize a Pioneer Neuron Scaffold to Steer Growing Thalamocortical Axons. Cerebral Cortex (New York, NY), 26(7), 3323-3334.
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6

Comprehensive Immunohistochemistry Panel

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Primary antibodies: rabbit anti-GFP (1:500, Invitrogen); sheep anti-GFP (1:1000, AbD Serotec); mouse anti-SV2 (1:100, Developmental Studies Hybridoma Bank); goat anti-ChAT (1:200, Millipore); guinea pig anti-VGluT2 (1:3000, Millipore); mouse anti-Cre (1:500, Millipore); goat anti-CTB (1:8000, List Biological Laboratories); mouse anti-CTB (1:500, Abcam); goat anti-WGA (1:500, Vector Labs); rabbit anti-tdTomato (1:1000, Clontech; Living Colors DsRed); and goat anti-DTR (1:400, Santa Cruz Biotechnology; HBEGF). Appropriate fluorophore-conjugated secondary antibodies were from the Jackson ImmunoReseach antibody series.
Azim E., Jiang J., Alstermark B, & Jessell T.M. (2014). Skilled reaching relies on a V2a propriospinal internal copy circuit. Nature, 508(7496), 357-363.
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7

Visualization of NTS Neuron Connectivity

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We used immunofluorescence multiple labeling to visualize VGLUT2+ and GAD65+ terminals and BDA3K+ neurons in NTS. The sections were rinsed and incubated in streptavidin Alexa-488 (1:200; Molecular Probes) for 24 hours at 4°C to label the BDA3K+ SSN-projecting neurons in NTS. The sections were then rinsed and incubated in mouse anti-GAD65 (Sigma-Alrdich, 1:200) and either guinea pig anti-VGLUT2 (Millipore, 1:1000) or rabbit anti-VGLUT2 (Sigma-Aldrich, 1:1000), for 24 hours at 4°C. The antibodies were diluted with 0.1M PB – 0.8% Triton X-100. After rinsing, the tissue was incubated in donkey anti-mouse IgG-Alexa-594 (1:200; Molecular Probes), and either goat anti-guinea pig IgG-Alexa-647 or donkey anti-rabbit IgG-Alexa-647 (1:200; Molecular Probes), overnight at 4°C. The sections were then mounted on gelatin-coated slides, air-dried, and cover-slipped in glycerol phosphate buffer (9:1). Sections were viewed and images captured using a Zeiss 710 confocal laser-scanning microscope.
Li C., Fitzgerald M.E., Del Mar N, & Reiner A. (2016). Disinhibition of Neurons of the Nucleus of Solitary Tract that Project to the Superior Salivatory Nucleus Causes Choroidal Vasodilation: Implications for Mechanisms Underlying Choroidal Baroregulation. Neuroscience letters, 633, 106-111.
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8

Immunohistochemistry Profiling of Neural Markers

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Immunohistochemistry was performed as previously described (Lammel et al., 2012). Briefly, animals were intracardially perfused with 4% paraformaldehyde in PBS and post-fixed at 4 °C overnight. 50 µm coronal sections were sliced on a Leica VT1000 vibratome. Primary antibodies used were: guinea pig anti-Vglut1 (1:250; Millipore), guinea pig anti-Vglut2 (1:1000; Millipore), rabbit anti-TH (1:1000; Millipore), rabbit anti-NeuN (1:2000; Abcam), mouse anti-GAD65 (1:250; DSHB U. of Iowa), guinea pig anti-substance P (1:500; Abcam). For spaghetti monster experiments, epitope antibodies used were: rabbit anti-FLAG (1:1000; Sigma), goat anti-myc (1:1000; Abcam), mouse anti-V5 (1:1000; Life Technologies), rat anti-HA (1:1000; Roche). Alexa Fluor dyes conjugated to either 405, 488, 568, or 647 were used for secondary antibodies (Life Technologies, Abcam). All dilutions used were 1:1000 except for Alexa Fluor 405 at 1:500. All images were acquired with an Olympus FluoView FV1200 confocal microscope.
Knowland D., Lilascharoen V., Pacia C.P., Shin S., Wang E.H, & Lim B.K. (2017). Distinct ventral pallidal neural populations mediate separate symptoms of depression. Cell, 170(2), 284-297.e18.
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9

Immunohistochemical Analysis of Synaptic Proteins

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Antibodies used in this study include rabbit anti-Synpo (1:400; Synaptic Systems, Goettingen, DE; RRID:AB_887825), guinea pig anti-Synpo (1:400; Synaptic Systems, RRID:AB_10549419), guinea pig anti-VGluT2 (1:500; Millipore, Burlington, MA, US; RRID:AB_1587626), rabbit anti-Synaptoporin (1:200, Proteintech, Rosemont, IL, US; RRID:AB_2878022). Anti-rabbit and anti-guinea pig secondary antibodies conjugated to Alexa Fluor 488 (1:400) were obtained from Thermo Fisher Scientific.
Speranza L., Filiz K.D., Goebel S., Perrone-Capano C., Pulcrano S., Volpicelli F, & Francesconi A. (2022). Combined DiI and Antibody Labeling Reveals Complex Dysgenesis of Hippocampal Dendritic Spines in a Mouse Model of Fragile X Syndrome. Biomedicines, 10(11), 2692.
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10

Immunofluorescence Antibody Staining Protocol

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Primary antibodies and dilutions used in this study: rabbit anti-Mef2c (1:500, Cell Signaling), mouse anti-mGluR2 (1:1500, Advanced Targeting Systems), mouse anti-NeuN (1:1000, Millipore), mouse anti-Calbindin (1:5000, Swant), goat anti-parvalbumin (1:2500, Swant), rat anti-GFP (1:1000, Nacalai Tesque), guinea pig anti-vGluT2 (1:2000, Millipore), rabbit anti-GFAP (1:500, Sigma Aldrich), and guinea pig anti-zebrin (1:1000, Frontier Institute). All secondary antibodies used were obtained from molecular probes and diluted 1:1000 prior to use.
Kamath S.P, & Chen A.I. (2018). Myocyte Enhancer Factor 2c Regulates Dendritic Complexity and Connectivity of Cerebellar Purkinje Cells. Molecular Neurobiology, 56(6), 4102-4119.
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