Mouse brain samples were extracted using the same method described above for the MS analysis. For each analysis, 5–20 μg of sample was added per lane, separated on 12% Bis-Tris Pre-Cast gels (Thermo Fisher) and transferred to PVDF membranes using 7.5% BSA in TBS for blocking. All primary antibodies were used overnight at 1:1000. Antibodies were used against TH (tyrosine hydroxylase; EMD Millipore, 657012 RRID:AB_696697 ), DAT (dopamine transporter; EMD Millipore, MAB369 RRID:AB_2190413 ), Ddc (aromatic-L-amino-acid decarboxylase; Abcam, ab3905 RRID:AB_304145 ), NSE (neuron specific enolase; Abcam, ab53025 RRID:AB_881756 ), PKC-β2 (protein kinase C beta-2; Abcam, ab32026 RRID:AB_779042 ), Akt (protein kinase B; Cell Signaling, 9272 RRID:AB_329827 ), Lmp7 (proteasome subunit beta type-8; Abcam, ab3329 RRID:AB_303708 ), and α-synuclein clone D37A6 (Cell Signaling Technology, #4179 RRID:AB_1904156 ) or clone Syn211 (Sigma-Aldrich Cat# S5566, RRID:AB_261518 ). Antigen-antibody complexes were detected using an Odyssey LC scanner (LiCor) after incubation with appropriate secondary antibodies. Densitometry was used to quantify intensity of protein bands.
Mab369
Manufactured by Abcam
MAB369 is a mouse monoclonal antibody that detects Connexin 43 (Cx43). Cx43 is a gap junction protein that plays a role in cell-to-cell communication. The antibody can be used in various applications, such as Western blotting, immunohistochemistry, and immunocytochemistry, to detect Cx43 expression in different cell types and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Ab54835, by Abcam (2 mentions)
Ab19353, by Abcam (2 mentions)
Fluoromount aqueous mounting medium, by Merck Group (2 mentions)
Trueblack, by Biotium (2 mentions)
Ab53025, by Abcam (1 mentions)
Ab3329, by Abcam (1 mentions)
Ab3905, by Abcam (1 mentions)
Bis tris precast gel, by Thermo Fisher Scientific (1 mentions)
Odyssey lc scanner, by LI COR (1 mentions)
Ab32026, by Abcam (1 mentions)
Bca assay kit, by Merck Group (1 mentions)
Tissue tearor, by Biospec (1 mentions)
Immobilon ecl, by Merck Group (1 mentions)
4 protocols using mab369
1
Proteomic Profiling of Mouse Brain Samples
2
Immunofluorescence Staining and Imaging Protocol
3
Multicolor Immunocytochemistry Protocol
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Cells were fixed with 4% paraformaldehyde for 20 min at 4 °C, permeabilized with 0.125% Triton X-100 in PBS and blocked with True Black (Biotium) according to the manufacturer’s directions and then 10% normal goat serum (NGS) for 1 h. Primary antibodies were applied overnight in 5% NGS at 1:1000. Secondary antibodies were applied for 1–2 h at room temperature, diluted 1:1000 in 5% NGS. Coverslips were then mounted in Fluoromount Aqueous Mounting Medium (Sigma). Antibodies used in this study include rabbit anti-GFP (Clontech, 612460), rat anti-DAT (Millipore MAB369), sheep anti-DBH (abcam ab19353), rabbit anti-RhoA (abcam, ab54835), rabbit anit-NeuN (Abcam, ab177487) and rabbit anti-EAAT3 (α-Diagnostics, EAAC11-A).
4
Striatal Protein Extraction and Western Analysis
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Protein extraction and Western analysis was performed as previously described (Janezic et al., 2013 (link)). Striatal tissue was homogenized in PBS (pH 7.4) containing 1% Igepal CA-630, 0.1% SDS, 0.5% sodium deoxycholate, and protease inhibitor mixture using a Tissue Tearor (Biospec Products, Inc.). Protein content was quantified using a BCA assay kit (Sigma) and proteins were analyzed by Western blotting under reducing, non-denaturing conditions. Primary antibodies used were: mouse anti-alpha synuclein 1:500 (Abcam ab1903) and rat anti-DAT 1:1,000 (Millipore; mab369) and anti-beta actin (HRP-conjugated) antibody (Abcam ab49900) at 1:50,000. Bands were visualized using horseradish peroxidase-conjugated goat anti-mouse or goat anti-rat IgG (Bio-Rad) and the chemiluminescent ECL+ kit (GE Healthcare) or immobilon ECL (Millipore). Bands were quantified using ImageJ software.
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