Sentence examples for stringent buffer from inspiring English sources

With ReCLIP, protein complexes are crosslinked in situ and then lysed in RIPA, a stringent buffer designed expressly to be compatible with antibody-antigen interactions while preventing nonspecific and/or weak interactions.

Under less stringent buffer conditions that retain more protein-protein contacts, one or more of the antibodies must now be unable to recognize their target.

We found that none of these KH domains bound their reported RNA ligands in SBB buffer (data not shown), which contains 200 m m salt and so is a moderately stringent buffer for protein RNA association.

In order to separate the cytoplasmic and nuclear fraction of the colon cancer cells they were first lysed using a less stringent Buffer A (10 mM HEPES pH7.9, 10 mM KCl pH7.0, 100 μM EDTA pH 8.0, 1 mM DTT, 0.5 mM PMSF) on ice for 15 minutes followed by centrifugation at 6800xg for 2 minutes.

Briefly, several cycles of washes were done initially with a nonstringent buffer (1 M NaCl, 67 mM NaH2PO4, 6.7 mM EDTA, 0.01% Tween 20) at 25°C and then with stringent buffer [100 mM MES, 0.1 M Na+, 0.01% Tween 20] at 50°C.

The IP materials were washed twice with stringent buffer (100 m m Tris-HCI, pH 7.4, 500 m m LiCI, 0.1% Triton X-100, 1 m m DTT, 2 μg mL−1 leupeptin, 2 μg mL−1 aprotinin, 1 m m phenylmethylsulfonyl fluoride), and twice with the IP buffer.

Briefly, we fractionated the samples by resolubilization in increasingly stringent buffers (Tris-buffered saline, 1% Triton X-100, 1% sarcosyl, 8 M urea) as previously described.

Samples were then washed with a series of stringent buffers to remove non-specifically bound DNA fragments.

Antibody-bound chromatin was precipitated with Protein G conjugated agarose beads, washed with gradient stringent buffers, and eluted with elution buffer as per the manufacturer's instructions.

Filters were washed at 65°C in increasingly stringent buffers (2 × sodium chloride/sodium citrate SSC, 0.1% SDS to 0.1× SSC, 0.1% SDS) until counts were approximately 1000 cpm.

After hybridization, the arrays were washed and stained in an Affymetrix wash station following Affymetrix Eukaryotic protocol, in a series of washes in non-stringent buffer (6× SSPE, 0.01% Tween-20) followed by stringent washes (100 mM MES, 0.1 M NaCl, 0.01% Tween-20).