Sentences with phrase «goat serum with»

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Samples were incubated with primary antibody (1/100 in PBST with BSA and goat serum) for 4 °C at 12 hours.

The cells were fixed with 4 % formaldehyde (10 min), permeabilized with 0.1 % Triton X-100 for 5 minutes and then blocked in 1 % BSA / 10 % normal goat serum / 0.3 M glycine in 0.1 % PBS - Tween for 1h.

The cells were incubated at 37 °C for 3h in media containing different concentrations of ab120223 (FK506) in DMSO, fixed with 100 % methanol for 5 minutes at -20 °C and blocked with PBS containing 10 % goat serum, 0.3 M glycine, 1 % BSA and 0.1 % tween for 2h at room temperature.

The cells were fixed with 4 % formaldehyde (10 min) and then blocked in 1 % BSA / 10 % normal goat serum / 0.3 M glycine in 0.1 % PBS - Tween for 1h.

Isolated retinas were blocked and permeabilized with retina blocking buffer (10 % normal goat serum, Dako, #X0907, in 0.5 % Triton - X 100 and 1 % BSA in PBS) for 1 h at RT..

Sections were incubated in 0.3 % hydrogen peroxide in PBS for 20 min, washed in PBS - T (0.01 M phosphate - buffered saline, 0.2 % Triton X-100) and blocked 1 h with 10 % normal goat serum (NGS) in PBS - T.

The cells were fixed with 4 % formaldehyde (10 min), permeabilized in 0.1 % Triton X-100 for 5 minutes and then blocked in 1 % BSA / 10 % normal goat serum / 0.3 M glycine in 0.1 % PBS - Tween for 1h.

Cells were fixed with 100 % methanol (5 min) at room temperature and incubated with PBS containing 10 % goat serum, 0.3 M glycine, 1 % BSA and 0.1 % triton for 1h at room temperature to permeabilise the cells and block non-specific protein - protein interactions.

Coverslips plated with acutely dissociated hNPCctx - GDNF or hNPCctx were fixed with 4 % paraformaldehyde, washed with PBS, blocked in 5 % normal donkey serum and 0.1 % Triton X-100, and incubated with goat anti-GDNF (1 ∶ 100; R&D Systems) primary antibody followed by donkey anti-goat Cy3 - conjugated secondary antibody (1 ∶ 1000; Jackson IR).

After blocking for 1 hour with 10 % normal goat serum (NGS), the slides were incubated with 200 μl primary antibody, either a polyclonal rabbit anti-Borrelia (Accurate chemicals, Westbury, NY) at 1:200 or the anti-OspA monoclonal, CB10, with a 1:30 dilution of hybridoma supernatant for 1 hour at room temperature (RT).

The cells were fixed with 100 % methanol for 5 minutes, permeabilized with 0.1 % Triton X-100 for 5 minutes and then blocked with 1 % BSA / 10 % normal goat serum / 0.3 M glycine in 0.1 % PBS - Tween for 1 hour.

The membranes were incubated with mouse sera obtained 3 - 5 weeks after inoculation (1:200 dilution), followed by three wash steps for 15 min each and incubation for 1 - 2 h with peroxidase - conjugated sheep anti-mouse IgG (whole molecule) or goat anti-mouse polyvalent immunoglobulins (1:10,000 dilution)(Sigma - Aldrich).

Approximately 10 million HUF1 cells were trypsinized through a 5 min exposure to 0.05 % trypsin - EDTA (Invitrogen), exposed to MEF media to inactivate the trypsin and then washed twice with ice cold PBS + 2 % goat serum (PBS - G).

For immunofluorescence, the cells were fixed in 4 % paraformaldehyde / PBS for 20 minutes, washed twice with PBS, and blocked with 4 % goat serum in PBS for 30 min, with all procedures performed at room temperature.

Fixed cells were blocked for one hour in 0.1 % Triton X-100 PBS supplemented with 10 % goat serum and 1 % BSA, followed by incubation with the primary antibody at 4 °C overnight in 0.1 % Triton X-100 with 8 % goat serum and 1 % BSA.

The cells were incubated at 37 °C for 24 hours in media containing different concentrations of ab120847 (metformin hydrochloride) in water, fixed with 4 % formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10 % goat serum, 0.3 M glycine, 1 % BSA and 0.1 % tween for 2 hours at room temperature.

The cells were incubated at 37 °C for 6h in media containing different concentrations of ab120723 (baicalein) in DMSO, fixed with 4 % formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10 % goat serum, 0.3 M glycine, 1 % BSA and 0.1 % tween for 2h at room temperature.

The concentration of in - situ antibody - AChR complexes was determined by precipitation with goat anti-rat serum in the presence of normal rat serum.