Anti-Tyrosine Hydroxylase Antibody (A104316)

Affinity purified Anti-Tyrosine Hydroxylase Antibody (A104316) conjugated with ALEXA647 reliably detects TH expressed in murine peripheral blood mononuclear cells (PBMC). Whole blood acquired from C57B6/J mice via cardiac puncture was diluted 1:1 in sterile PBS and layered on top of Ficoll-Histopaque (GE Healthcare, 17-1440-03) and centrifuged at 400g, room temperature, for 20 minutes with acceleration set to minimum and brakes off. Cells acquired from interface of Ficoll and whole blood were washed twice in sterile PBS, and immediately fixed on ice using the eBioscience Fixation/Permeablization kit (eBioscience, 88-8824-00) for 30 minutes, then washed twice with 1mL permeabilization buffer. Intracellular staining for TH was accomplished by addition of 2µL of affinity purified Anti-Tyrosine Hydroxylase Antibody (A104316) directly conjugated to Alexa647 in 100uL staining volume (1:50 dilution from stock antibody conjugate solution at 2.2mg/mL) and incubated at room temperature for 30 minutes. Following two washes in 1mL permeabilization buffer, data were immediately acquired on a Sony SP6800 Spectral Analyzer, and analyzed in a FCS Express 7 software (DeNovo).