rabbit antiserum rabbit igg, universal negative control serum  (Biocare Medical)

Journal: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology

Article Title: Dopamine Increases CD14 + CD16 + Monocyte Transmigration Across the Blood Brain Barrier: Implications for Substance Abuse and HIV Neuropathogenesis

doi: 10.1007/s11481-017-9726-9

Figure Lengend Snippet: CD14+CD16+ monocytes express greater surface D1R and D5R, and less D4R, than T cells. Mature monocytes and T cells from independent donors were incubated with antibodies to D1R, D5R, D4R or negative control rabbit serum or goat IgG. After incubation in primary antibody, cells were incubated with PE-labeled anti-rabbit or anti-goat IgG secondary antibody. Stained cells were analyzed by flow cytometry and the MFI was calculated by subtracting the appropriate negative control antibody or serum staining from the staining obtained with each dopamine receptor antibody. Significance was determined by two-tailed unpaired Mann-Whitney test. **p<0.01 (D5R on mature monocytes, n=17, compared to T cells, n=9); ***p<0.001 (D1R on mature monocytes, n=19, compared to T cells, n=10) (D4R on mature monocytes, n=18, compared to T cells, n=9).

Article Snippet: For dopamine receptor staining, mature monocytes and purified, unactivated T cells (1×10 6 per tube) were stained with antibodies to dopamine D1 receptor (D1R) (324390), dopamine D3 receptor (D3R) (324402), dopamine D5 receptor (D5R) (324408) (1:5 dilution) (Calbiochem/EMD Millipore, Billerica, MA), dopamine D2 receptor (D2R) (B-10 and H-50) (1:10 dilution), dopamine D4 receptor (D4R) (N-20) (1:10 dilution) (Santa Cruz Biotechnology, Santa Cruz, CA) or negative control rabbit serum, rabbit IgG, mouse IgG2a (Sigma-Aldrich, St. Louis, MO) or goat IgG (Santa Cruz Biotechnology) in 50 ul on ice for 30 minutes.

Techniques: Incubation, Negative Control, Labeling, Staining, Flow Cytometry, Two Tailed Test, MANN-WHITNEY