- PA5-27569 - Invitrogen Antibodies SLC32A1 antibody

Submitted references Precise Coordination of Three-Dimensional Rotational Kinematics by Ventral Tegmental Area GABAergic Neurons.

Hughes RN, Watson GDR, Petter EA, Kim N, Bakhurin KI, Yin HH
Current biology : CB 2019 Oct 7;29(19):3244-3255.e4

Dose-dependent reversal of KCC2 hypofunction and phenobarbital-resistant neonatal seizures by ANA12.

Carter BM, Sullivan BJ, Landers JR, Kadam SD
Scientific reports 2018 Aug 10;8(1):11987

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of VGAT using 50 µg of mouse brain lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a VGAT polyclonal antibody (Product # PA5-27569) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of VGAT using 30 µg of U87-MG lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a VGAT polyclonal antibody (Product # PA5-27569) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of VGAT was performed by separating 30 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a VGAT Polyclonal Antibody (Product # PA5-27569) at a dilution of 1:5000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using VGAT Polyclonal Antibody (Product # PA5-27569). Non-transfected (–) and transfected (+) 293T whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with VGAT Polyclonal Antibody (Product # PA5-27569) diluted at 1:2,500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using VGAT Polyclonal Antibody (Product # PA5-27569). Non-transfected (–) and transfected (+) 293T whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with VGAT Polyclonal Antibody (Product # PA5-27569) diluted at 1:5,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-VGAT Polyclonal Antibody (Product # PA5-27569) and band at ~ 50 kDa corresponding to VGAT was observed across the panel tested except in Mouse and Rat Ovary and Rat Liver which are reported to be negative for VGAT. Tissue extracts (30 µg lysate) of Mouse Retina (Lane 1), Mouse Cerebellum (Lane 2), Mouse Liver (Lane 3), Mouse Ovary (Lane 4), Rat Retina (Lane 5), Rat Cerebellum (Lane 6), Rat Liver (Lane 7) and Rat Ovary (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:5000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of VGAT was performed in Cultured rat E18 primary cortical neuron, DIV 8. Cells fixed in 4% paraformaldehyde at RT for 15 min. Green: VGAT Polyclonal Antibody (Product # PA5-27569) diluted at 1:250. Red: beta Tubulin 3/ TUJ1, stained by beta Tubulin 3/ TUJ1 antibody. Blue: Fluoroshield with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: VGAT Polyclonal Antibody detects VGAT protein by immunofluorescent analysis. Sample: DIV9 rat cortical neuron and Glia cell cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: VGAT stained by VGAT Polyclonal Antibody (Product # PA5-27569) diluted at 1:250. Red: Tau, a Axon marker, stained by Phospho-Tau (Ser262) Polyclonal Antibody [GT287]diluted at 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry (Frozen) analysis of VGAT was performed in frozen sectioned adult mouse retina tissue using VGAT Polyclonal Antibody (Product # PA5-27569) at a dilution of 1:250 (Green). Red: beta Tubulin 3/ TUJ1, stained by beta Tubulin 3/ TUJ1 antibody diluted at 1:250. Blue: Fluoroshield with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry (Frozen) analysis of VGAT was performed in frozen-sectioned adult mouse cerebellum tissue using VGAT Polyclonal Antibody (Product # PA5-27569) at a dilution of 1:250 (Green). Red: beta Tubulin 3/ TUJ1, stained by beta Tubulin 3/ TUJ1 antibody diluted at 1:500. Blue: Fluoroshield with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry (Paraffin) analysis of VGAT was performed in paraffin-Embedded adult mouse retina tissue using Green: CRABP2 Polyclonal Antibody (Product # PA5-27451) at a dilution of 1:250. Red: beta Tubulin 3/ TUJ1, stained by beta Tubulin 3/ TUJ1 antibody diluted at 1:500. Blue: Fluoroshield with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: VGAT Polyclonal Antibody detects VGAT protein at cytoplasm by immunohistochemical analysis. Sample: Paraffin-embedded mouse eye. Green: VGAT stained by VGAT Polyclonal Antibody (Product # PA5-27569) diluted at 1:250. Red: beta Tubulin 3/ Tuj1, a cytoskeleton marker, stained by beta Tubulin 3/ Tuj1 antibody [GT11710] diluted at 1:500. Blue: Fluoroshield with DAPI. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of paraffin-embedded CL1-5 xenograft, using VGAT (Product # PA5-27569) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 vGLUT1 vs. vGAT expression 24 h post-ischemia. ( A ) Representative Western blots showing vGLUT1 and vGAT expression at P7. The blots presented for each group were cropped from separate gels for improvement in clarity and conciseness of presentation. Vertical solid black lines delineate the separate blots between groups, while horizontal dotted black lines delineate separate proteins run on different gels for the same sample. All gels were run in the same experimental conditions (see methods for details). (Full-length blots of each tested presented in Supplementary Figure 7 ). ( B ) Quantification of Western blots shown in A. vGLUT1 is downregulated by ischemia and rescued by all doses of ANA12 + PB in a dose-dependent manner. vGAT is not modulated by ischemia. ( C ) Representative Western blots showing vGLUT1 and vGAT expression at P10. ( D ) Quantification of blots shown in C. vGLUT1 is downregulated by ischemia and rescued in a dose-dependent manner by ANA12 + PB. vGAT is unaffected by ischemia. Significance to PB alone: * P < 0.05, ** P < 0.01, *** P < 0.001. Significance to naive control: # P < 0.05, ## P < 0.01, ### P < 0.001. Significance to 5 mg/kg of ANA12 + PB: @ P < 0.05, @@ P < 0.01, @@@ P < 0.001. Ipsilateral to contralateral: gamma P < 0.05, gammagamma P < 0.01, gammagammagamma P < 0.001.

PA5-27569

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