Anti-Dopamine Transporter antibody [6V-23-23]

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [6V-23-23] (AB128848)

ab128848 staining of dopamine transporter in mouse brain (substantia nigra) tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/500) for two hours at room temperature. A Biotin-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody.

Carl Hobbs (Kings College, London, United Kingdom

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Transporter antibody [6V-23-23] (AB128848)

IHC image of Dopamine Transporter staining in a section of formalin-fixed paraffin-embedded mouse brain performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab128848, 5ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody. 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

Ap

Western blot - Anti-Dopamine Transporter antibody [6V-23-23] (AB128848)

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 ab128848 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.

Rat cerebellum (lane 3) was included as a negative control. This tissue does not express dopamine transporter protein.

All lanes:

Western blot - Anti-Dopamine Transporter antibody [6V-23-23] (ab128848) at 5 µg/mL

Lane 1:

Forebrain (Mouse) Tissue Lysate at 20 µg

Lane 2:

Forebrain (Rat) Tissue Lysate at 20 µg

Lane 3:

Cerebellum (Rat) Tissue Lysate at 20 µg

Lane 4:

Brain (Rat) Striatal Tissue Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/10000 dilution

Predicted band size: 68 kDa

Observed band size: 18 kDa,69 kDa

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Exposure time: 20min