A second and third set of tissues was assayed for D1 and D2. For D1, tissues were incubated in primary antibody rat anti-D1 (1:1,000, Sigma Aldrich D2944) for 48 hours at 4°C. Specificity of D1 antibody was validated by omission of primary antibody and by a previous prebsorption and Western blot study (24 (link)). Tissues were incubated for 1 hour in donkey anti-rat/Alexa 488 (1:1,000, Life Technologies A21208) and counterstained with DAPI. For D2, tissues underwent antigen retrieval for 30 minutes in 10 mM sodium citrate buffer (pH 8.5) to unmask immunosignal and were incubated in primary antibody mouse anti-D2 (1:200, Santa Cruz sc-5303) for 48 hours at 4°C. Specificity of D2 antibody was validated by omission of primary antibody and by previous preabsorption and Western blot studies (25 (link), 26 (link)). Tissues were incubated for 1 hour in donkey anti-mouse/Alexa 488 (1:1,000, Life Technologies A21202).
Protocol detail
Immunohistochemical Detection of Dopamine Receptors
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Anti antibody
Antibody
Antibody specificity
Antigen
Buffer
Dapi
Donkey
Mouse
Sodium citrate
Tissues
Western blot
Corresponding Organization : University of Portland
Top 5 similar protocols
Variable analysis
independent variables
- Incubation of tissues in primary antibody rat anti-D1 (1:1,000, Sigma Aldrich D2944) for 48 hours at 4°C
- Incubation of tissues in primary antibody mouse anti-D2 (1:200, Santa Cruz sc-5303) for 48 hours at 4°C
- D1 and D2 expression
- Incubation of tissues in donkey anti-rat/Alexa 488 (1:1,000, Life Technologies A21208) for 1 hour
- Incubation of tissues in donkey anti-mouse/Alexa 488 (1:1,000, Life Technologies A21202) for 1 hour
- Counterstaining of tissues with DAPI
- Antigen retrieval of tissues for 30 minutes in 10 mM sodium citrate buffer (pH 8.5)
- Positive control: Omission of primary antibody for D1 and D2
- Positive control: Previous preabsorption and Western blot studies for D1 (24) and D2 (25, 26)
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!