ICC: PLP fixed rat brain sections. The antiserum has been found to stain specific cells in various regions, including the Medial septum, Nucleus accumbens, Dentate gyrus, Globus pallidus, Medial forebrain bundle, Cortex regions 1-3, Substantia Nigra reticulata, and the Ventral tegmental area.
WB: whole rat brain homogenate.
Lyophilised:Reconstitute with 0.1ml of PBS which contains 10 mg/ml BSA.
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 49 kDa (predicted molecular weight: 49 kDa).Can be blocked with Dopamine Receptor D1 peptide (ab137968). With this antibody, we have found that blocking with 5% goat or donkey serum significantly reduces background as compared to BSA or milk.
Dopamine receptor whose activity is mediated by G proteins which activate adenylyl cyclase.
Detected in caudate, nucleus accumbens and in the olfactory tubercle.
Belongs to the G-protein coupled receptor 1 family.
Cell membrane. Endoplasmic reticulum membrane. Transport from the endoplasmic reticulum to the cell surface is regulated by interaction with DNAJC14.
ab40653 (1:500) staining Dopamine Receptor D1 in human cerebellum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of cytoplasmic and membrane regions in the fibrous glial cells. Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
Western blot - Dopamine Receptor D1 antibody (ab40653)
All lanes : Anti-Dopamine Receptor D1 antibody (ab40653) at 1/1000 dilution
Lane 1 : Human brain tissue lysate - total protein (ab29466) Lane 2 : Rat Cortex Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 49 kDa Observed band size : 49 kDa Additional bands at : 42 kDa,75 kDa. We are unsure as to the identity of these extra bands.
Cheng HW et al. Identification of thioridazine, an antipsychotic drug, as an antiglioblastoma and anticancer stem cell agent using public gene expression data. Cell Death Dis6:e1753 (2015).
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