Sentences with phrase «lysing for»

Cells were washed with ice - cold PBS, lysed for 15 minutes on ice with M - PER protein extraction reagent (Pierce), scraped, and spun at 14,000 rpm for 10 minutes at 4 °C.

Not exact matches

EGFR / WT and KO cells were cultured overnight and starved for 24 h. Cells were treated with capsaicin (50 μmol / L) for 30 min and then TPA (20 ng / mL) for 2 h. Cells were lysed and cox - 2 mRNA levels were analyzed by RT - PCR.

EGFR / WT and KO MEFs were treated with capsaicin (0, 10, or 50 μmol / L) for 30 min followed by TPA (20 ng / mL) for 4 h. Cells were lysed and COX - 2 expression analyzed by Western blot.

Cells were washed once with PBS and lysed in 0.02 mM NaOH for 16 hr at 4 °C, and then stored at — 20 °C until analysis.

For Western blot analysis, 106 BaF3 cells were starved for 4 h and lysed in 250 μL of Laemmli buffer (BioRaFor Western blot analysis, 106 BaF3 cells were starved for 4 h and lysed in 250 μL of Laemmli buffer (BioRafor 4 h and lysed in 250 μL of Laemmli buffer (BioRad).

Cell supernatants were discarded and cell pellets were lysed of RBCs and then used for staining with antibodies for flow cytometry analysis.

Cells were then lysed and probed for transcription of Nrf2, Keap1, or Bach1 using the QuantiGene II RNA plex (Affymetrix).

For this protocol, samples were lysed in 1 × RIPK2 lysis buffer [50 mM Tris (pH 7.5), 0 mM MgCl2, 1 % Triton X-100, 1 mM dithiothreitol, 1 mM EDTA, 1 mM EGTA, with freshly added 1 mM β - glycerophosphate and protease inhibitors (phenylmethylsulfonyl fluoride and aprotinin)-RSB- and immunoprecipitated overnight using 1.5 μg rabbit anti-RIPK2 antibody.

(E) 106 BaF3 cells stably transduced with JAK2 V617F, Y931C and V617F / Y931C double JAK2 mutant were treated for 30 min with 300 nM CMP6 inhibitor or with DMSO as a control (− condition), lysed and subjected to Western blot analysis.

Cultured cells were lysed in radioimmunoprecipitation assay (RIPA) buffer (50 μl RIPA per 1 × 106 cells; 50 mM Tris / HCl; 150 mM NaCl, 1 % Triton X-100, 1 mM EDTA, 1 mM NaF, 1 mM Na3VO4, 1 μg / ml leupeptin and 1 μg / ml pepstatin) for 1 h on ice, and insoluble debris was removed by centrifugation.

(A) 106 BaF3 Aut (V658I), Aut (F958V) and BaF3 cells stably transduced with M - RAS or (B) JAK1 V658I and F958V mutants were treated for 30 min with 500 nM CMP6 inhibitor or with 0.1 % DMSO as a control (− condition), lysed and subjected to Western blot analysis.

Isolation of cells from outer, middle and inner colony zones was performed under microscopic guidance (Figure S4), after which cells were immediately lysed and prepared for Q - RTPCR.

48 hours after transfection, cells were lysed with 100 µL / well 1 × passive lysis buffer (PLB, Promega) for 15 minutes with shaking.

Confluent 10 cm plates were fixed with 1 % PFA at room temperature for 10 minutes, lysed in SDS lysis buffer (50 mM Tris, 10 mM EDTA, 1 % SDS, Roche Complete protease inhibitors, pH 8.1), scraped and collected into 1.5 mL microcentrifuge tubes, and DNA was sonicated to 200 — 800 bp fragments with a Branson Sonifier 250 set to 30 % power / 90 % duty, four 10 second pulses.

For quantification of phagocytosis, alveolar macrophages were infected at an MOI of 20 for 1 h, then media was removed, extracellular bacteria were killed with gentamycin (200 μg / ml for 10 min at 37 C), and macrophages were lysed with 0.2 % triton X-100 (10 min at room temperaturFor quantification of phagocytosis, alveolar macrophages were infected at an MOI of 20 for 1 h, then media was removed, extracellular bacteria were killed with gentamycin (200 μg / ml for 10 min at 37 C), and macrophages were lysed with 0.2 % triton X-100 (10 min at room temperaturfor 1 h, then media was removed, extracellular bacteria were killed with gentamycin (200 μg / ml for 10 min at 37 C), and macrophages were lysed with 0.2 % triton X-100 (10 min at room temperaturfor 10 min at 37 C), and macrophages were lysed with 0.2 % triton X-100 (10 min at room temperature).

WT and Lpar2 - / - T cells were incubated with 1 μM LPA for the periods indicated, and were lysed immediately thereafter.

Essentially, this acts as a method of «pre-digesting» the protein by separating (i.e. lysing) peptide bonds; hence the time for digestion and absorption of amino acids will be reduced.

Supplies Serratia peptidase enzyme to help protect sensitive GI tissues from irritation and lysozyme for its unique lysing action on bacterial and yeast cell walls

Along with new voices from the Stockholm 89plus Open Call: Philip Berlim & Louise Dahl, Elis Burrau, Arvida Byström & Maja Malou Lyse, The Council for Grand Art, Natalia Marina Fuentes Araya, George Nordmark, Rojda Sekerös and Klara Ström.

A high affinity binding site for HP1 can be produced by silencing Lys9 of histone H3 by methylation with mammalian histone lysing methyltransferase, a suppressor of variegation 39H1 (SUV39H1).

MONTREAL — Lyse Lemieux, the first woman to be named chief justice of the Quebec Superior Court, announced today that she is retiring after being cited for impaired driving.

«Lyse Lemieux has served the administration of justice with skill and dignity for over two decades.

The authors thank Hélène Beauchesne and Lyse Desmarais - Gervais for project administration and data collection coordination, and Muriel Rorive and Nathalie Fréchette for data management.