Sentence examples for buffer before incubation with from inspiring English sources

Protein blots were incubated in blocking buffer before incubation with primary antibody according to the manufacturers' protocols.

Elution from the beads is achieved in an acidic reaction buffer before incubation with the cofactor.

Cells were then permeabilized with 1% Triton X-100 in PBS (pH 7.4) and blocked with 5% bovine serum albumin in PBS buffer, before incubation with the DAPI and secondary fluorescent antibody goat anti-human IgG.

Tissue slides were microwaved in 0.01  M sodium citrate buffer (pH 6) near boiling for 20 min, and cooled for 30 min in the buffer before incubation with the primary antibodies.

The primary antibody was removed and sections were washed for 3×20 min in blocking buffer before incubation with the secondary antibody and Alexa-Fluor-phalloidin for 2 hrs at RT.

Sections were then treated with 3% hydrogen peroxidase (Sigma-Aldrich) for 10 min, and boiled in 10 m m citrate buffer before incubation with primary antibody [Pax6: diluted 1: 300; Covance, Princeton NJ, USA; Emx2: 1: 200; Sigma-Aldrich; in 0.3% PBS with 0.3% Triton X-100 (PBS-T) and 3% horse serum; Vector Laboratories, Peterborough, England].

Non-specific sites were blocked with SuperBlock blocking buffer (Pierce Biotechnology) before incubation with secondary antibodies.

In subsequent experiments the supernatants were first preincubated with a polyclonal anti-factor H antibody (50 μg ml−1) and/or with the GB24 anti-MCP (25 μg ml−1) mAb (30 min, 37°C) or buffer (control) before incubation with I-C3b and factor I (90 min, 37°C).

The mononuclear cells in the buffy layer were collected and washed with Phosphate Buffer Saline (PBS) before incubation with TH for 24 hours at increasing concentrations.

To prepare whole-cell extracts, cells at 90% confluent were washed in phosphate-buffered saline (PBS) before incubation with lysis buffer (1% Triton X-100, 150 mM NaCl, 10 mM Tris, pH 7.4, 1 mM EDTA, 1 mM EGTA, pH 8.0, 0.2 mM Na3VO4, 0.2 mM phenylmethylsulfonyl fluoride and 0.5% Nonidet P-40) on ice for 10 min.

The membranes were immersed at room temperature in phosphate-buffered saline supplemented with 0.2% Tween 20 and 5% nonfat dry milk (blocking buffer) for 1 h before incubation with primary serum (diluted 1 1,000 in blocking buffer) for 1 h.