dc cy5  (Jena Bioscience)

Journal: bioRxiv

Article Title: Transcription swells chromosomes in vitro

doi: 10.1101/2024.09.25.614905

Figure Lengend Snippet: a) A DNA molecule of the lambda bacteriophage was labeled on specific sites with fluorophores (dC-Cy5, purple star) for imaging and with biotins for surface immobilization in an open microfluidic chip. The surface-immobilized lambda DNA molecule (yellow dashed line) was stretched by flow (yellow solid-line circle). b) A representative fluorescence time-lapse montage shows a DNA molecule before, during, and after a moderate flow pulse. We tracked the position of the fluorescent dye to measure DNA extension, as shown for three different displacement examples. c) The DNA extension by flow was measured after incubating the DNA molecules for 30 minutes in cell-free expression systems with active transcription (Tx), without Tx by adding 500 nM rifampicin, and in phosphate-buffered saline (PBS) solution as a protein-free reference. The data were combined for Tx(ON), Tx(OFF), and buffer from n=3,2,3 independent experiments with mean values as yellow bars and individual molecules as circles.

Article Snippet: The purified lambda DNA was further nick-translated with 25 µM dG, 25 µM dA, 25 µM dT, 15 µM dC, 10 µM dC-Cy5 (Jena Biosciences) and cleaned as described above.

Techniques: Labeling, Imaging, Lambda DNA Preparation, Fluorescence, Expressing, Saline

Journal: International Journal of Nanomedicine

Article Title: Engineered hepatitis B virus surface antigen L protein particles for in vivo active targeting of splenic dendritic cells

doi: 10.2147/IJN.S32813

Figure Lengend Snippet: Flow cytometric analysis of ex vivo attachment of α-DC-ZZ-BNCs complexes to splenic DCs. ( A ) Accumulation of Cy5-labeled α-DC-ZZ-BNC complexes to isolated splenic DCs. Splenic DCs were incubated with each Cy5-labeled α-DC-ZZ-BNC complex and subjected to flow cytometric analysis. Fractions of DCs were pre-defined by the forward scatter/side scatter dot plots derived from CD11c+ cells. Distributions of Cy5-derived fluorescence in ZZ-BNC-incubated DCs and untreated DCs are indicated as closed and open histograms, respectively. Antibodies against DCs are shown in the upper left of each panel. The percentages (%) of ZZ-BNC+ cells in DCs are indicated as numbers. ( B ) Mean fluorescent intensities of the Cy5-labeled α-DC-ZZ-BNC complexes in DCs. Notes: Mean fluorescent intensity derived from DCs incubated with Cy5-labeled BNCs was defined as 100. Measurements were performed in triplicate. Error bars represent the SD. Abbreviations: DC, dendritic cell; ZZ-BNC, BNC displaying ZZ domains; DC, dendritic cell; SD, standard deviation.

Article Snippet: DCs (2.5–5.0 × 10 5 cells) were mixed with each Cy5-labeled α-DC-ZZ-BNC complex (1 μg as ZZ-L protein), incubated at 4°C for 30 minutes, and then analyzed with the flow cytometer BD FACScan Canto II (BD Biosciences, San Jose, CA) with linear amplification for forward/side scatter and logarithmic amplification for FITC and Cy5 fluorescence.

Techniques: Ex Vivo, Labeling, Isolation, Incubation, Derivative Assay, Fluorescence, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Engineered hepatitis B virus surface antigen L protein particles for in vivo active targeting of splenic dendritic cells

doi: 10.2147/IJN.S32813

Figure Lengend Snippet: Flow cytometric analysis of in vivo attachment of α-DC-ZZ-BNC complexes to splenic DCs. ( A ) Distributions of Cy5-derived fluorescence in splenic DCs isolated from Cy5-labeled α-DC-ZZ-BNC complex-injected mice and untreated mice are indicated by closed and open histograms, respectively. ( B ) Distributions of Cy5-derived fluorescence in splenic CD11c − cells isolated from Cy5-labeled α-DC-ZZ-BNC complex-injected mice. Notes: The percentages (%) of ZZ-BNC + cells in DCs and CD11c − cells are indicated as numbers. Abbreviations: DC, dendritic cell; ZZ-BNC, BNC displaying ZZ domains.

Article Snippet: DCs (2.5–5.0 × 10 5 cells) were mixed with each Cy5-labeled α-DC-ZZ-BNC complex (1 μg as ZZ-L protein), incubated at 4°C for 30 minutes, and then analyzed with the flow cytometer BD FACScan Canto II (BD Biosciences, San Jose, CA) with linear amplification for forward/side scatter and logarithmic amplification for FITC and Cy5 fluorescence.

Techniques: In Vivo, Derivative Assay, Fluorescence, Isolation, Labeling, Injection

Journal: International Journal of Nanomedicine

Article Title: Engineered hepatitis B virus surface antigen L protein particles for in vivo active targeting of splenic dendritic cells

doi: 10.2147/IJN.S32813

Figure Lengend Snippet: Incorporation of Cy5-labeled α-CD11c (clone N418)-ZZ-BNC complexes by splenic DCs isolated from Cy5-labeled α-DC-ZZ-BNC complex-injected mice. Notes: Z-stack projections of splenic DCs were generated from deconvolved slices using the maximum intensity criteria. Fluorescence derived from ZZ-BNCs and CD11c molecules is indicated in red and green, respectively. Scale bars, 5 μm. Abbreviations: DC, dendritic cell; ZZ-BNC, BNC displaying ZZ domains; DIC, differential interference contrast.

Article Snippet: DCs (2.5–5.0 × 10 5 cells) were mixed with each Cy5-labeled α-DC-ZZ-BNC complex (1 μg as ZZ-L protein), incubated at 4°C for 30 minutes, and then analyzed with the flow cytometer BD FACScan Canto II (BD Biosciences, San Jose, CA) with linear amplification for forward/side scatter and logarithmic amplification for FITC and Cy5 fluorescence.

Techniques: Labeling, Isolation, Injection, Generated, Fluorescence, Derivative Assay