Heroin (First Pharmaceutical Factory of Shenyang, Shenyang, China), the selective dopamine D1 receptor agonist SKF-38393, D1 receptor antagonist SCH-23390, and D3 receptor agonist PD-128907 (Sigma-Aldrich) were dissolved in 0.9% saline to obtain the required final concentration. The doses were based on previous reports (Cote and Kuzhikandathil, 2014 (link); Sheng et al., 2015 (link)). Rabbit polyclonal antibodies against dopamine D1 (ab20066), D5 (ab40656), D2 (ab21218), D3 (ab42114), and D4 (ab135978) receptor were purchased from Abcam Technology. The mouse monoclonal antibodies against GAPDH and the horseradish peroxidase-conjugated anti-rabbit or anti-mouse secondary antibodies were purchased from Santa Cruz Technology.
Ab40656
Manufactured by Abcam
Ab40656 is a laboratory equipment product. It is designed for general laboratory use. The core function of this product is to [description not available].
Automatically generated - may contain errors
Lab products found in correlation
Ab20066, by Abcam (2 mentions)
Ab21218, by Abcam (1 mentions)
Skf38393, by Merck Group (1 mentions)
Pd 128907, by Merck Group (1 mentions)
Sch23390, by Merck Group (1 mentions)
Ab52476, by Abcam (1 mentions)
Ab45689, by Abcam (1 mentions)
Ab85367, by Abcam (1 mentions)
Ab42114, by Abcam (1 mentions)
Ab211265, by Abcam (1 mentions)
2 protocols using ab40656
1
Dopamine Receptor Pharmacology in Heroin Use
2
Dopamine Receptor Protein Expression
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The proteins were extracted from the striatum tissues using radioimmunoprecipitation assay (RIPA) lysis buffer (Ukzybiotech, Beijing, China) according to the manufacturer's instructions. The protein concentration in the lysates was evaluated using a BCA Protein Assay Kit. Proteins were then separated on an SDS-PAGE and transferred to a polyvinylidene difluoride membrane. Membranes were blocked with 5% BSA-TBST and then incubated with DA1R antibody (ab20066, 1 : 1000), DA2R antibody (ab85367, 1 : 1000), DA3R antibody (ab42114, 1 : 1000), DA4R antibody (ab20424, 1 : 500), DA5R antibody (ab40656, 1 : 200), PKA antibody (ab211265, 1 : 1000), CAM (ab45689, 1 : 1000), and CAMKII (ab52476, 1 : 1000) (Abcam) overnight at 4°C. Then, the blots were washed, incubated with 5% BSA-TBST, and washed again. Consequently, samples were incubated with horseradish peroxidase-conjugated secondary antibody at room temperature for 2 h. Band detection was performed using the enhanced chemiluminescence (ECL) detection kit. The intensity of the detected bands was calculated densitometrically using the Gel Image system ver.4.00 (Tanon, China).
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