Journal: Biophysical Journal

Article Title: Improved Model of Fluorescence Recovery Expands the Application of Multiphoton Fluorescence Recovery after Photobleaching in Vivo

doi: 10.1016/j.bpj.2009.04.020

Figure Lengend Snippet: Comparison of in vitro experimental data with flow, fit to the diffusion-only and diffusion-convection models. A series of experimental fluorescence recovery curves for FITC-BSA and FITC-2000 kDa dextran were taken over a wide range of known flow

Article Snippet: For in vitro testing of the flow model, fluorescent samples were produced by mixing fluorescein isothiocyanate (FITC) conjugated to bovine serum albumin (BSA) or 2000 kDa dextran (dextran) (Molecular Probes/Invitrogen) diluted to 1 mg/mL in phosphate buffered saline (PBS) with 1 μ L/mL red fluorescent microspheres (FluoSpheres; Molecular Probes/Invitrogen).

Techniques: In Vitro, Flow Cytometry, Diffusion-based Assay, Convection, Fluorescence

Journal: Biophysical Journal

Article Title: Improved Model of Fluorescence Recovery Expands the Application of Multiphoton Fluorescence Recovery after Photobleaching in Vivo

doi: 10.1016/j.bpj.2009.04.020

Figure Lengend Snippet: Results of fitting in vitro experimental data with flow to the diffusion-convection model. A series of experimental fluorescence recovery curves for FITC-BSA and FITC-2000 kDa dextran were taken over a wide range of known flow speeds (plotted

Article Snippet: For in vitro testing of the flow model, fluorescent samples were produced by mixing fluorescein isothiocyanate (FITC) conjugated to bovine serum albumin (BSA) or 2000 kDa dextran (dextran) (Molecular Probes/Invitrogen) diluted to 1 mg/mL in phosphate buffered saline (PBS) with 1 μ L/mL red fluorescent microspheres (FluoSpheres; Molecular Probes/Invitrogen).

Techniques: In Vitro, Flow Cytometry, Diffusion-based Assay, Convection, Fluorescence

Journal: Glycobiology

Article Title: Glycoproteomics enabled by tagging sialic acid- or galactose-terminated glycans

doi: 10.1093/glycob/cws144

Figure Lengend Snippet: GAL efficiently labels Gal and GalNAc residues on cell surface glycoproteins. ( A ) Asialo-K20 cells were incubated in PBS with 5 mg/mL BSA, pH 6.7, containing 250 μM aminooxy-biotin in the presence or absence of 10 mM aniline and 50 U/mL galactose

Article Snippet: For the oxime ligation step, cells were suspended to 1 × 107 cells/mL in PBS/5% BSA, pH 6.7 containing 250 μM aminooxy-biotin, aminooxy-AF488 (Invitrogen Corporation) or aminooxy-FLAG peptide and 10 mM aniline (Sigma–Aldrich) for 90 min at 4°C with end-over-end mixing.

Techniques: Incubation

Journal: The Journal of Biological Chemistry

Article Title: TDP2/TTRAP Is the Major 5?-Tyrosyl DNA Phosphodiesterase Activity in Vertebrate Cells and Is Critical for Cellular Resistance to Topoisomerase II-induced DNA Damage *

doi: 10.1074/jbc.M110.181016

Figure Lengend Snippet: The 5′-TDP activity of His-TDP2 prefers DSBs and liberates ligatable 5′ termini from DNA 5′-phosphotyrosine termini. A , purified recombinant human His-TDP2 (100 n m ) was incubated with 50 n m duplex substrate harboring a DSB with a 5′-phosphotyrosine ( P-Y ) terminus ( top ) in the absence or presence of the indicated concentrations of orthovanadate for 1 h at 37 °C and reaction products separated by denaturing PAGE and detected by autoradiography. Right , quantification of the fraction (%) of total labeled oligonucleotide converted to 5′-phosphate (19-mer) product is shown. Data are the average (± range) of two independent experiments. B , purified recombinant human His-TDP2 (100 n m ) was incubated with duplex substrate as above in the absence or presence of the indicated concentrations of BSA or phosphotyrosine-BSA. C , DNA substrates with a 5′-phosphotyrosine terminus present in single-stranded DNA ( top ), in a blunt-ended DNA duplex ( middle ), or in a nicked duplex ( bottom ), were incubated with the indicated concentrations of His-TDP2 for 1 h at 37 °C and reaction products analyzed as above. The middle graph is the quantification (% of total labeled oligonucleotide converted to 5′-P 19-mer product) of the above experiment, from three independent experiments mean (±S.E.). Asterisk denotes statistically significant difference ( p

Article Snippet: Reactions were carried out by incubating the indicated substrate (50 nm ) with whole cell protein extract (250 ng/ml) or the indicated concentration of recombinant human His-TDP2/TTRAP protein, which was expressed in Escherichia coli and prepared as described previously , in 25 mm HEPES, pH 8.0, 130 mm KCl, 10 mm MgCl2 , 1 mm DTT and in the presence or absence of orthovanadate, BSA, or phosphotyrosine-BSA (Sigma) as indicated at 37 °C for 60 min.

Techniques: Activity Assay, Purification, Recombinant, Incubation, Polyacrylamide Gel Electrophoresis, Autoradiography, Labeling