Sentence examples for buffer and subsequently stained from inspiring English sources

Remaining tissue aggregates were digested to single-cell suspension by incubation at 37°C with 0.01% DNase I and 0.15% collagenase D. Cells from the suspension were washed in cold FACS buffer and subsequently stained with anti-MTS24 antibody and sorted using a FACScalibur II sorter.

Permeabilized cells were incubated with PE-conjugated TCRβ antibody and phospho S6 antibody for 45 min at room temperature, washed in saponin buffer and subsequently stained with FITC conjugated donkey anti rabbit IgG (Jackson Immunoresearch, West Grove PA).

The PCR products together with a 100 bp DNA ladder (Invitrogen™) were separated by electrophoresis in 1.5% agarose gel in TAE buffer, and subsequently stained with ethidium bromide for visualization in GelDoc 2000 equipment (Bio-Rad™).

The cells were washed with ice-cold phosphate-buffered saline (PBS; Sigma; cat. no. D8537), resuspended in Annexin V-binding buffer, and subsequently stained with AnnexinV-fluorescein isothiocyanate and PI (BD Pharmingen, San Jose, CA, USA), according to the manufacturer's recommendation (Società Chimici Italiani; cat. no. IK-11120).

Twelve hours after transfection of 25 nmol/L STAT3 decoy or scramble ODN, cells were detached, washed and resuspended in 100 μl Annexin V binding buffer, and subsequently stained with Annexin V-FITC and propidum iodide (Sigma) according to the manufacturer's recommendation.

Fixation buffer was removed with two washes with permeabilization buffer (BD Pharmingen) and samples were split and subsequently stained for intracellular cytokines using 1/200 anti-IFN-γ-allophycocyanin, anti-IL-4-PE, anti-IL-10-allophycocyanin, anti-IL-13-allophycocyanin, anti-IL-17-PE, or the relevant isotype control for 20 min in Perm buffer.

After the indicated treatments, cells were washed and subsequently stained with 100  μL of 1x binding buffer containing 5  μL of Annexin V-FITC for 15 min at room temperature in the dark.

Gels were run in a Tris-Glycine SDS buffer system at 40 mA per gel for approximatively 7 h and subsequently stained with Coomassie PAGE Blue (Fermentas, St. Leon-Rot, Germany) according to the manufacturer's instructions.

Five milliliters of each eluate were pipetted onto fibroblast cultures, incubated, and subsequently stained.

Cells were subsequently fixed in paraformaldeyde at defined time points and subsequently stained with cristal violet.

After washing with a staining buffer, cells were incubated with anti-mouse CD16/32 (Fc block, e-Bioscience) for 30 min on ice to block the FcR non-specific binding and subsequently stained for surface markers with PE-anti-CD4, FITC-anti-CD3ε, PerCy-anti-CD8 or isotype controls for 30 min on ice.