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After washes in blocking buffer, the blots were incubated simultaneously in 2nd Abs linked to Cy3 and Cy5.

After 5 times 5 min washes in blocking buffer, the blots were incubated for 1 hr in 2nd Ab linked to alkaline phosphatase (AP) diluted 1∶10,000 in blocking buffer.

After being blocked with 5% nonfat milk powder in TBS buffer, the blots were probed with the antibodies specific for the indicated proteins.

After extensive washing with TBST buffer, the blots were then incubated with goat anti-rabbit, horseradish peroxidase-conjugated secondary antibody for 1 h at room temperature.

After rinsing with the washing buffer, the blots were incubated with the secondary antibodies (either horseradish peroxidase-conjugated goat anti-rabbit IgG or horseradish peroxidase-conjugated goat anti-mouse IgG; 1 2000; Dingguo Biotechnology) at room temperature for 45 min.

Blots were incubated in blocking buffer (p38α MAPKp44/42 MAPK (Erk1/2), After four washes with buffer, the blots were incubated with corresponding peroxidase-conjugated secondary antibody in corresponding blocking buffer for 2 h at room temperature.

The supernatant was mixed with 5X loading buffer and the blots were run using the NuPage Novex 4 12% gradient Bis-Tris gels (Invitrogen) with MOPS running buffer (Invitrogen).

After prehybridization at 68°C for 1 hour in Ultrahyb Ultrasensitive Hybridizaion Buffer (Ambion), the blots were subjected to hybridization with Digoxigenin-labeled RNA probes overnight at 68°C.

After applying a stripping buffer (Thermo Fisher), the blots were re-probed with antibodies against GAPDH (Santa Cruz Biotechnology) or total Erk1/2 (Cell Signaling).

Excess radioactive probe was washed out with 2x SSC, 0.1% SDS buffer, and the blots were exposed for one week to phosphorimaging screen (Fujifilm) and scanned with Fujifilm FLA5100 reader (Fujifilm Co., Ltd., Tokyo, Japan).

Biotinylated oligonucleotides (see Additional file 1, table S1), designed using FastPCR [ 41], were then used for hybridization in Northern Max buffer (Ambion), and the blots were developed by chemiluminescent detection using the BrightStar Biodetect kit (Ambion).