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Our Abpromise guarantee covers the use of ab20424 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500. Detects a band of approximately 48-53 kDa (predicted molecular weight: 48 kDa).|
|IHC-P||Use at an assay dependent concentration. PubMed: 22155235|
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat gastric tissue labelling Dopamine Receptor D4 with ab20424 at 1/1000. Dopamine Receptor D4 was not found in the gastric mucosa (e - left) but was observed in the gastric enteric plexus (f - right). Paraffin sections were baked at 60°C for 2 hours, then deparaffinized in xylene for 20 minutes. Slides were rehydrated in descending concentrations of alcohol and transferred from 70% alcohol solution to distilled water for rinsing. Before staining, slides were pretreated with 10 mM citric acid (pH 6.0) at 92–98°C for 10 min and incubated in 10% normal serum and 0.1% Triton X-100 for 30 min. The sections were then incubated with the primary antibody, followed by the appropriate secondary antibody. Sections were washed in PBS and mounted in 90% glycerin PBS.
ab20424 stained PC12 cells. The cells were 4% formaldhyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab20424 at 1/100) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
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