Sentences with phrase «containing lysate»

Following the provided protocol, a portion of the RNA - containing lysate (5 — 10 µl) was hybridized at 54 degrees C overnight to RNA specific magnetic capture beads in the presence of blocking buffers, proteinase K and preordered mRNA probe sets specific for the genes of interest: HMOX1, Nrf2, Keap1, Bach1, and GAPDH.

Not exact matches

Subsequently, lungs were digested with DMEM containing 1.25 mM CaCl2, 200 U ml − 1 collagenase I and 10 μg ml − 1 DNase I at 37 °C for 1 h. Single - cell suspensions of the digested organ were prepared by passing through 18G and 19G cannula syringes and filtering the lysates through a 100 μm cell strainer.

METHODS: We treated 3T3 - L1 adipocytes with 2.5 mmol / l R (+) alpha - lipoic acid for 2 to 60 min, followed by assays of: 2 - deoxyglucose uptake; glucose transporter 1 and 4 (GLUT1 and GLUT4) subcellular localization; tyrosine phosphorylation of the insulin receptor or of the insulin receptor substrate - 1 in cell lysates; association of phosphatidylinositol 3 - kinase activity with immunoprecipitates of proteins containing phosphotyrosine or of insulin receptor substrate - 1 using a in vitro kinase assay; association of the p85 subunit of phosphatidylinositol 3 - kinase with phosphotyrosine proteins or with insulin receptor substrate - 1; and in vitro activity of immunoprecipitated Akt1.

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