Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: ELISA-based analysis for reactivity of anti-PspA hkR36A MAbs with recombinant PspAs representing the 6 PspA clades a

Article Snippet: Six- to 8-week-old female BALB/c mice were immunized subcutaneously with 25 μg recombinant PspA R36A emulsified with Imject alum (1:1 [wt/wt]) (Pierce, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant

Journal: eLife

Article Title: CTP and parS coordinate ParB partition complex dynamics and ParA-ATPase activation for ParABS-mediated DNA partitioning

doi: 10.7554/eLife.65651

Figure Lengend Snippet: Apparent disassembly or exchange rate constants (min −1 ) and ParB F /ParA F ratio from fits of TIRFM wash and FRAP experiments. The apparent dissociation (or FRAP) rate constants ( k off ) were obtained for individual time-trajectories by single-exponential curve fitting (except ** where the rate of the faster decay, (68 ± 0.5%) of a double-exponential fit is shown), and the mean and SEM for the set of independent experiments are shown (except * where standard deviation among non-independent repeats within an experiment is shown). (N is the number of separate experiments, with total number of binding/wash cycles for repeated data collection in parenthesis.) ParA:ParB ratios were calculated from the final phase of the individual association time-trajectories (except *** where it was based on the beginning part of the washing phase in the presence of ParB F 1-42 R36A ), and the mean and SEM for the set of independent experiments are shown in italics. N.D., not done.

Article Snippet: ParB F 1-42 and ParB F 1-42 R36A were synthesized de novo (Genscript).

Techniques: Standard Deviation, Binding Assay

Journal: eLife

Article Title: CTP and parS coordinate ParB partition complex dynamics and ParA-ATPase activation for ParABS-mediated DNA partitioning

doi: 10.7554/eLife.65651

Figure Lengend Snippet: ATPase fit parameters. ATPase measurements were performed with ParA F (1 μM) and different mutants of ParB F , 60 μg/ml EcoRI-digested pBR322 DNA plus Scram- or parS F -DNA fragment and CTP or CDP, as indicated in the column headings. Assays were repeated ‘ N ’ times, each data set of an assay was fit after subtraction of background measured without ParA F to a modified Hill equation: v − v 0 = (v max [B] n ) / (K A n + [B] n ), and the mean and standard error of the mean (SEM) of the fit parameters for the N measurements are shown. For [B] on the x-axis, total ParB F concentration was used instead of free ParB F concentration due to technical issues in estimating the free ParB F concentration and the meanings of K A and the cooperativity factor (n) here differ from those in the standard adaptation of the Hill equation. v max is the maximum stimulated ParA F ATPase turnover rate, K A is the apparent total concentration of ParB F necessary for half maximum stimulation, and n is the apparent cooperativity coefficient.

Article Snippet: ParB F 1-42 and ParB F 1-42 R36A were synthesized de novo (Genscript).

Techniques: Modification, Concentration Assay

Journal: eLife

Article Title: CTP and parS coordinate ParB partition complex dynamics and ParA-ATPase activation for ParABS-mediated DNA partitioning

doi: 10.7554/eLife.65651

Figure Lengend Snippet: ( A ) ParB F 1-42 R36A -mCherry (10 μM) and ParA F -eGFP (1 μM) preincubated with ATPγS were infused into the nsDNA-carpeted flow cell and then ( B ) washed with buffer containing ATPγS. ( C ) The washing experiment of B was repeated with buffer containing ATPγS and ParB F 1-42 R36A -mCherry (10 μM). ( D ) ParA F -ATPase activity was measured in the presence of EcoRI-digested pBR322 DNA (60 μg/ml) as a function of ParB F 1-42 R36A concentration. See legend and and for additional details.

Article Snippet: ParB F 1-42 and ParB F 1-42 R36A were synthesized de novo (Genscript).

Techniques: Activity Assay, Concentration Assay