Sentence examples for blocking solution was subsequently from inspiring English sources
To prevent non-specific staining, fixed cells were first blocked in 10% natural rabbit serum (DakoCytomation) for 30 min at RT. Blocking solution was subsequently removed and primary antibodies were added at the following dilutions: anti-ERK5, 1 : 100; anti-Mcm2, 1 : 200.
Secondary antibody (rabbit anti-sheep HRP diluted to 200 μg·mL−1 in block or sheep anti-mouse HRP diluted to 400 μg·mL−1 in blocking solution) was then incubated for 60 min and subsequently washed four times in PBS.
Subsequently, non-specific binding was blocked with 10% normal goat serum in 0.05% Tween in PBS (PBST) for 30 min at RT after which the blocking solution was replaced by primary antibody solution diluted in 2% normal goat serum (NGS) in PBST (60 min at RT).
Blocking solution was removed and primary antibody in blocking solution applied at the required dilution.
Blocking solution was used to block the endogenous peroxidase activity.
For membrane blocking, a 5% Top-Block solution was used.
The solutions were subsequently filtered.
Membranes were then washed with blocking solution and proteins were subsequently detected by chemiluminescence (Amersham Pharmacia Biotech, NJ, USA).
After a second treatment with blocking solution containing mouse serum and avidin sections were subsequently stained with a biotinylated anti-CD45.2 primary (eBioscience, 104) antibody and streptavidin-DyLight549 (Vector Laboratories).
These membranes were subsequently incubated with blocking solution, exposed to the DNA probe specified below and repeatedly washed using a standard Southern hybridization procedure.
