Anti-VGluT1 antibody [N28-9]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VGluT1 antibody [N28-9] (AB134283)

IHC image of VGluT1 staining in rat brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab134283, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• WB

Project

Western blot - Anti-VGluT1 antibody [N28-9] (AB134283)

This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab134283 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-VGluT1 antibody [N28-9] (ab134283) at 5 µg/mL

Lane 1:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 20 µg

Lane 2:

Brain (Rat) Tissue Lysate at 20 µg

Secondary

All lanes:

Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/10000 dilution

Predicted band size: 61 kDa

Observed band size: 50 kDa,55 kDa

true

Exposure time: 20min

• WB

Lab

Western blot - Anti-VGluT1 antibody [N28-9] (AB134283)

This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab134283 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-VGluT1 antibody [N28-9] (ab134283) at 5 µg

Lane 1:

Human Brain Tissue Lysate at 20 µg

Lane 2:

Rat Brain Tissue Lysate at 20 µg

Lane 3:

Mouse Brain Tissue Lysate at 20 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/5000 dilution

Predicted band size: 61 kDa

Observed band size: 200 kDa,50 kDa,55 kDa

true

Exposure time: 30s

• WB

CiteAb

Western blot - Anti-VGluT1 antibody [N28-9] (AB134283)

VGluT1 western blot using anti-VGluT1 antibody [N28-9] ab134283. Publication image and figure legend from Torres, L., Robinson, S. A., et al., 2018, J Neuroinflammation, PubMed 29471842.

ab134283 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab134283 please see the product overview.

T. gondii infection decreased VGLUT2 expression in the mouse olfactory bulb. a Brains from mock-infected control mice and mice orally infected with 10 ME49 cysts for 30 and 60 days were collected after perfusion with ice-cold 4% PFA. Brain sections were fixed and stained with anti-VGLUT1 (green), anti-VGLUT2 (red), and mounted with DAPI (blue). Whole brain images were scanned using Leica Biosystems Scanscope and processed with image scope. Representative images were taken from the olfactory bulb at × 20 magnification. n = 4–5 mice per group, the experiment was repeated three times. Scale bar = 50 μm. b, c At least five different fields of the olfactory bulb and pre-frontal cortex were randomly selected from 2 to 4 mice per group. The intensity of VGLUT2 (b) and VGLUT1 (c) signal at different time points post infection was quantified. d Representative images of the olfactory bulb of mock-infected and T. gondii-infected mice stained with anti-GAD67 (green) and VGLUT2 (red). Scale bar = 50 μm, n = 4 mice per group. e The mean fluorescent intensity for GAD67 (green) was measured using Axiovision software from five different fields (n = 4 mice per group). f Representative image from the olfactory bulb of infected mice stained with anti-GAD67 (green) and anti-T. gondii (red). Scale bar = 50 μm. g Western blot analysis of VGLUT1/2 (55 and 65 kDa, respectively) and GAD67 (67 kDa) protein levels using whole brain samples from mock-infected controls and T. gondii-infected mice at days 30 and 60 post infection. GAPDH (37 kDa) was used as loading control. Blot shown is representative of three independent experiments. h–j Densitometric analysis of Western blot data from g. The results are expressed as the ratio of densities of GAD67 or VGLUT1/2-specific bands to GAPDH bands. Data are shown as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, **** p < 0.0001

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