D-5 Dopamine Receptor Antibody – Rabbit Polyclonal

Immunocytochemistry: This antiserum has been found to stain specific cells in various regions of PLP fixed rat brain sections at a dilution of 1:2500. This includes cortex layer 5 and the parafascicular nucleus.

Western Blot: Western immunoblots using whole rat brain homogenate resulted in a single band being detected at ~49 kD at 1:200 dilution.

Western Blot Protocol: 1. After SDS-PAGE on 12% gels and electrophoretic transfer to PVDF membrane. block the membrane overnight with 4% normal goat serum in TBS/Tween-20 buffer.
2. Wash x 2 with TBS/Tween-20.
3. Apply the rabbit polyclonal antibody after dilution to at least 1:200 (Note: higher dilutions may be needed). Use 2% normal goat serum in TBS/Tween-20 as buffer for the primary antibody. Let the primary antibody bind for 2-4 hours.
4. Wash x 3 with TBS/Tween-20.
5. Apply affinity purified HRP-goat anti-rabbit IgG antiserum diluted 1:2500 (dilution may vary depending upon supplier) in 2% normal goat serum in TBS/Tween-20. Incubate 1-2 hours. Note: greater sensitivity may be achieved using ABC techniques.
6. Wash x 4 for 5 minute/wash with TBS/Tween-20.
7. Develop color using the enhanced DAB reaction.