PMC:4307275 / 18760-19937 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/4307275","sourcedb":"PMC","sourceid":"4307275","source_url":"https://www.ncbi.nlm.nih.gov/pmc/4307275","text":"Optimization of the amount of DNA for FCA-based analysis using a KR-sGFP reporter system. The fluorescent signals exceeding the gating threshold were detected in single live protoplasts. (A) The sGFP fluorescence was detected using the FITC-A channel of the flow cytometer and its values were normalized by the fluorescent intensity of the protoplasts transfected using 0 μg DNA. The results are presented in arbitrary units. The transfection ratio was calculated by the proportion of transfected protoplasts relative to total protoplasts gated by side scattering and forward scattering procedure on FCA. The transfected protoplasts are determined by their fluorescence intensity exceeding the threshold value (103) on FCA; The results are normalized by transfection ratio of the protoplasts transfected using 2 μg DNA and presented in arbitrary units and (B) The expression efficiency of proteins was compared using western blot analysis. Polyclonal rabbit antibody against sGFP was used at a titer of 1:5000 and the image of PAGE gel was used to show relative quantities of the loaded proteins. The graph shows the relative band intensities quantified using LAS4000 software.","tracks":[]}