Sentence examples for milk block from inspiring English sources
After at least 1 h, the milk block was removed, and 100 μL primary antibody diluted into the NFDM added for 1 h at 37 °C.
After 1 h defatted milk block, the membrane was incubated with the anti-GABAB1 antibody (Abcam, 1 : 1000) and anti-GABAB2 antibody (Santa Cruz; 1 : 200) and then with horseradish peroxidase- (HRP-) conjugated secondary antibody (1 : 5000).
Membranes were blocked for 1 h in Milk block buffer (1% 10 m Tris/HCl (pH7.4), 150 m NaCl, 4% fat free milk powder), and were then incubated with primary antibody on a rotator at 4°C overnight.
Protein was separated on either NuPAGE 4 12% gels (Invitrogen) using the MES buffer system, or by conventional SDS PAGE using 14% polyacrylamide gels and transferred to either nitrocellulose (PrP blots, 3% BSA block) or PVDF (Sho blots, 5% non-fat skim milk block).
Again the plates were washed with PBST, incubated at room temperature for 1 h with 50 μL of secondary antibody [goat anti-human IgG (heavy and light chains) alkaline phosphatase-conjugate (AP), Bio-Rad 170 6521] diluted 1 1000 in 1% milk block.
After a series of washes in Milk block buffer and Tween buffer (1% 10 m Tris/HCl (pH7.4), 150 m NaCl and 0.2% Tween-20) membranes were incubated with HRP-conjugated sheep anti-mouse secondary antibody (Sigma-Aldrich, 1 : 1000) for 1 h followed by further washes and distilled water rinse.
Following a wash in phosphate buffered saline (PBS), the membranes were blocked in milk blocking buffer (ThermoPierce) for 1 hour at room temperature.
For dot blots, protein samples (1 4 μg) were directly applied to nitrocellulose membranes and incubated in 5 % milk blocking solution in 1 % Tween in Tris-base buffer saline (TTBS) for 60 min and then with 17G1 and J591 (dilution 1 1000) in PBS for 2 h at room temperature.
Polyclonal anti-chicken agrin antibody was diluted 1∶2000 in 5% milk blocking solution.
The membrane was then incubated in 3% milk blocking solution for 2 h at room temperature.
Unspecific binding was blocked by incubation in 5% milk blocking buffer (1× PBS, 5% nonfat milk and 0.1% Tween 20).
