Sentences with phrase «cell lysis buffer»

Cell pellets were resuspended in 1 ml cell lysis buffer [5 mM Pipes pH 8.0, 85 mM KCl, 0.5 % NP - 40] containing protease inhibitors and incubated for 10 min on ice.

Reactions were terminated with 1 ml of ice - cold PBS, and cells were centrifuged and resuspended in 50 μl of cell lysis buffer containing 1 % NP - 40 and protease inhibitors.

Not exact matches

Peripheral blood mononuclear cells were isolated by centrifugation with 96 % Ficoll (BD), followed by erythrocyte lysis with ACK lysis buffer.

At 48 hours, cells were washed twice with PBS and lysed in 1 × passive lysis buffer (Dual - Luciferase Reporter Assay, Promega).

Lane 1: HEK cells Lane 2: hfRPE cells Lane 3: hfRetina PVDF membrane RIPA lysis buffer with mixed phosphatase and protease inhibitors

Spleens were processed into cell suspensions through a 70 - μm strainer by a syringe plunger in RPMI 1640 with 10 % FBS and then treated with ACK lysis buffer to remove RBCs.

Lane 1: HEK cells Lane 2: hfRPE cells Lane 3: hfRetina Lane 4: Mouse liver Lane 5: Mouse RPE Lane 6: Mouse retina Lane 7: Zebrafish liver Lane 8: Zebrafish retina PVDF membrane RIPA lysis buffer with mixed phosphatase and protease inhibitor cocktail

Cells were lysed on ice using lysis buffer (10 mM Tris - HCl pH 7.4, 150 mM NaCl, 1 % Triton X-100, protease inhibitor mix G (Serva) and 2 mM Na3VO4).

Cells were lysed in RIPA lysis buffer.

For the coimmunoprecipitation experiments, HuH - 7 cells transfected with pcDNA - SRα / FLAG - Sp 1 were harvested and cell lysates were prepared using immunoprecipitation lysis buffer [20 mmol / L Tris - Cl (pH 7.5), 150 mmol / L NaCl, 10 % glycerol, and 1 % Triton X-100].

NHLF cells transfected with siRNA molecules were lysed in High Salt ELB lysis buffer [1 M Tris pH 8.0, 1 % NP - 40, 250 mM NaCl, 5 mM EDTA] supplemented with protease and phosphatase inhibitors (1x G - Biosciences Protease Arrest, 200 µM Na3VO4, and 1 mM PMSF).

Cells were lysed with TN1 lysis buffer (50 mM Tris, 125 mM NaCl, 1 % Triton X-100, 10 mM EDTA, 10 mM sodium fluoride and 10 mM sodium pyrophosphate) supplemented with protease inhibitor cocktail (1.2 mM AEBSF, 0.46 μM aprotinin, 14 μM bestatin, 12.3 μM E-64, 112 μM leupeptin, 1.16 μM pepstatin)(Amresco; Solon, OH)-RRB- and 1 mM sodium orthovanadate (Enzo Life Sciences; Farmingdale, NY).

Make up «direct lysis buffer» by adding 1 mL of Proteinase K (e.g. Sigma P4850) to 100 mL DirectPCR Lysis Reagent (cell)(Viagen Biotech, Cat # 302 lysis buffer» by adding 1 mL of Proteinase K (e.g. Sigma P4850) to 100 mL DirectPCR Lysis Reagent (cell)(Viagen Biotech, Cat # 302 Lysis Reagent (cell)(Viagen Biotech, Cat # 302 - C).

At the end of the desired treatment times, cell lysates were prepared in lysis buffer (1 % NP - 40, 0.5 mM Tris - HCl (pH 7.5), 0.14 M NaCl, 5 mM KCl, 5 mM EDTA, and 1 mM phenylmethylsulfonyl fluoride) plus serine / threonine phosphatase inhibitor cocktail.

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

Each single cell was carried through two rinses of PBS using a finely drawn glass capillary pipette under a dissecting microscope and then transferred in a minimal volume (0.5 µL) to a PCR tube containing lysis buffer, protease, tRNA and poly - T gripNA ™ probe (mTRAP ™ midi - kit, Active Motif, cat.