Ab111468

Manufactured by Abcam

Sourced in United Kingdom

Ab111468 is a recombinant monoclonal antibody. It is produced in mammalian cells and is designed to detect the target protein through immunoassays.

Automatically generated - may contain errors

Lab products found in correlation

Ab42114, by Abcam (2 mentions) Ab21218, by Abcam (2 mentions) Anti syp, by Santa Cruz Biotechnology (1 mentions) Anti snap25, by Santa Cruz Biotechnology (1 mentions) Ab20066, by Abcam (1 mentions) Ab85367, by Abcam (1 mentions) Anti munc18, by Synaptic Systems (1 mentions) Sc 13937, by Santa Cruz Biotechnology (1 mentions) Anti syntaxin, by Santa Cruz Biotechnology (1 mentions) Pargyline, by Merck Group (1 mentions) Ab9485, by Abcam (1 mentions) Horseradish peroxidase conjugated anti rabbit igg secondary antibody, by Abcam (1 mentions) Ab81296, by Abcam (1 mentions) Ab78021, by Abcam (1 mentions) Ab6721, by Abcam (1 mentions) Clarity western ecl substrate, by Bio-Rad (1 mentions)

5 protocols using ab111468

Western Blot Analysis of Neurotransmitter Receptors

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Immunoblot analysis was carried out under standard conditions (Marrocco et al., 2012 (link)) using the following primary antibodies: anti-tyrosine hydroxylase (TH H126, Santa Cruz, 1:1000); anti-D1 receptors (Ab20066, Abcam, 1:500), anti-D2 receptors (Ab85367, Abcam, 1:500), anti-DAT (high affinity DA transporter) (ab111468, Abcam, 1:1000); anti-adenosine receptors (sc-13937, Santa Cruz, 1:500), synaptic vesicle-associated proteins: anti-Rab3a (#107111, Synaptic Systems, 1:2000); anti-Munc18 (#116011, Synaptic Systems 1:2000; anti-SNAP 25 (sc-136,267, Santa Cruz 1:5000); anti-SYP (sc-9116 Santa Cruz 1:8000), and anti-syntaxin (sc-13994, Santa Cruz 1:4000). Secondary antibodies directed against rabbit or mouse antibodies (Amersham) were used at 1:7500 dilution. After immunoblotting, digitized images of bands immunoreactive to the target antibodies and actin were acquired (FUSION®) and the area of immunoreactivity corresponding to each band was measured using ImageJ imaging software. The ratios of the targets to actin were then determined and these values were compared to check statistical significance.

Marrocco J., Verhaeghe R., Bucci D., Di Menna L., Traficante A., Bouwalerh H., Van Camp G., Ghiglieri V., Picconi B., Calabresi P., Ravasi L., Cisani F., Bagheri F., Pittaluga A., Bruno V., Battaglia G., Morley-Fletcher S., Nicoletti F, & Maccari S. (2020). Maternal stress programs accelerated aging of the basal ganglia motor system in offspring. Neurobiology of Stress, 13, 100265.

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Immunocytochemical Analysis of Dopamine Transporter

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Nerve growth factor beta (NGF-β) and pargyline were obtained from Sigma-Aldrich (St Louis, MO, USA). Among the primary antibodies, the mouse monoclonal anti-β-Actin (AC15, A5141) was obtained from Sigma-Aldrich, and rabbit polyclonal anti-dopamine transporter (ab111468) from Abcam (Cambridge, UK). Secondary antibodies labeled with fluorescent dyes Alexa Fluor 488 goat anti-rabbit IgG antibody (A11034) and Alexa Fluor 594 goat anti-mouse IgG antibody (A11005) were purchased from Thermo Fisher (Waltham, MA, USA).

Haas A.J., Le Page Y., Zhadobov M., Sauleau R., Dréan Y.L, & Saligaut C. (2017). Effect of acute millimeter wave exposure on dopamine metabolism of NGF-treated PC12 cells. Journal of Radiation Research, 58(4), 439-445.

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Immunoblot Analysis of Synaptic Proteins

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The following antibodies were used according to the instructions supplied by the manufacturer: mouse anti-PSD95 (124011, Synaptic Systems), mouse anti-SYP (sc-55507, Santa Cruz Biotechnology), rabbit anti-NMDAR1 (ab32915, Abcam), mouse anti-VGLUT1 (135311, Synaptic Systems), rabbit anti-GAPDH (ab9485, Abcam), rabbit anti-DAT1 (ab111468, Abcam) and mouse anti-huDISC1 (3D4, Korth lab; Ottis et al., 2011 (link)). Immunoblot data were normalized to corresponding whole-lane densitometric volumes of protein-stained membranes (Welinder and Ekblad, 2011 (link)). Immunoblotting conditions were as previously described (Sialana et al., 2016 (link)) and antibody dilutions are provided in the Supplementary Table S1.

Sialana F.J., Wang A.L., Fazari B., Kristofova M., Smidak R., Trossbach S.V., Korth C., Huston J.P., de Souza Silva M.A, & Lubec G. (2018). Quantitative Proteomics of Synaptosomal Fractions in a Rat Overexpressing Human DISC1 Gene Indicates Profound Synaptic Dysregulation in the Dorsal Striatum. Frontiers in Molecular Neuroscience, 11, 26.

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Quantification of Dopamine Receptor Expression

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Equal amounts of proteins (70 μg) from all groups were loaded in the wells and the RCs were separated on 5–13% of blue native PAGE gels and the Western blot procedure was carried out using the procedure described previously (Kang et al., 2009 (link)). The details of antibodies used are as follows; D1R (1:5000, Abcam-ab81296, Cambridge, UK), D2R (1:5000, Abcam-ab21218, Cambridge, UK), D3R (1:5000, Abcam-ab42114, Cambridge, UK), pDAT (1:5000, DAT Thr53, Phosphosolutions-p435-53, Aurora, CO, USA) and DAT (1:5000, Abcam- ab111468, Cambridge, UK) and detected with horseradish peroxidase-conjugated anti-rabbit IgG secondary antibody (1:10000, Abcam-ab6721, Cambridge, UK). Immunoreactive bands were quantified by the software Image J (NIH). Coomassie blue R-350 stained membranes were used as loading control and normalized with the Western blot densitometric values (Welinder and Ekblad, 2011 (link); Sase et al., 2012 (link)).

Aher Y.D., Subramaniyan S., Shanmugasundaram B., Sase A., Saroja S.R., Holy M., Höger H., Beryozkina T., Sitte H.H., Leban J.J, & Lubec G. (2016). A Novel Heterocyclic Compound CE-104 Enhances Spatial Working Memory in the Radial Arm Maze in Rats and Modulates the Dopaminergic System. Frontiers in Behavioral Neuroscience, 10, 20.

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Western Blot Analysis of Dopamine Receptors

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Membrane proteins were transferred from BN-PAGE to PVDF membranes. After blocking of membranes for 1 h with 10 % non-fat dry milk in 0.1% TBST (100 mM Tris–HCL, 150 mM NaCl, pH 7.5, 0.1% Tween 20), membranes were incubated with primary antibodies D1R (diluted 1:5000, Abcam-ab78021, Cambridge, UK), D2R (diluted 1:5000, Abcam-ab21218, Cambridge, UK), D3R (diluted 1:5000, Abcam- ab42114, Cambridge, UK), pDAT (diluted 1:5000, DAT Thr⁵³, Phosphosolutions-p435-53, Aurora, United States) and DAT (diluted 1:5000, Abcam- ab111468, Cambridge, UK) and detected with horseradish peroxidase-conjugated anti-rabbit IgG (diluted 1:10000, Abcam- ab6721, Cambridge, UK). Membranes were developed with the Bio-Rad Clarity^TM Western ECL Substrate. Arbitrary optical densities of immunoreactive bands were measured by the Image J software program¹. After developing, total protein staining was done on PVDF membranes as previously described for loading control.

Karabacak Y., Sase S., Aher Y.D., Sase A., Saroja S.R., Cicvaric A., Höger H., Berger M., Bakulev V., Sitte H.H., Leban J., Monje F.J, & Lubec G. (2015). The effect of modafinil on the rat dopamine transporter and dopamine receptors D1–D3 paralleling cognitive enhancement in the radial arm maze. Frontiers in Behavioral Neuroscience, 9, 215.

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