Goat anti rabbit igg h l hrp 2 ab

Prior to staining, the endogenous peroxidase activity of cells was quenched by 3% H₂O₂ solution. Cells were first blocked with 2% BSA/0.1% casein in 1X PBS for 30 minutes. Rabbit anti-Lamin A IgG (LOT: L1293, Sigma-Aldrich) or anti-HSP90 IgG (LOT: SAB4300541, Sigma-Aldrich) diluted in PBS buffer containing 6% BSA was added to the cells. After 1-hour incubation, cells were washed three times (5 minutes each) with PBS containing 2% BSA, followed by another 1-hour incubation with goat anti-rabbit IgG (H+L) HRP-2’Ab (LOT: RA230590, ThermoFisher). Unbound antibodies were washed away with PBS with 2% BSA (5 min X 3), and fresh enzyme substrate (dopamine or DAB) was added to cells for 15 minutes incubation. The ideal staining result is strong chromogen signal of interested target locations with low nonspecific signals in background. To characterize the staining stability after storage, the stained cells were stored in 1X PBS at 4 °C, and washed with fresh PBS every four days. Images were captured every three weeks on the same cell subset with the same exposure and gain. For immunofluorescence imaging with QDs, after the PDA development step, amine-functionalized PEG-coated QDs (10 nM) were incubated with cells for 1 hour.