Journal: Frontiers in Plant Science

Article Title: Time-Resolved Investigation of Molecular Components Involved in the Induction of NO 3 – High Affinity Transport System in Maize Roots

doi: 10.3389/fpls.2016.01657

Figure Lengend Snippet: Western blot analysis carried out on the microsomal fraction extracted from the root tissue of maize seedlings treated with NO 3 – for the indicated times. (A) Western blot analysis performed using anti-PM H + -ATPase (upper panel), anti-NRT2.1 (middle panel) or anti-NRT3.1A (lower panel) antibodies. The same amount of proteins was loaded in each lane. The protein molecular weights were estimated using the ECL TM Plex Fluorescent Rainbow Markers (GE Healthcare Life Sciences). (B) Densitometric analysis of PM H + -ATPase, NRT2.1, and NRT3.1A. Data are expressed as ratio of the corresponding 0 h sample (signals were quantified using Quantity One software, Bio-Rad). Data are the means ± SE, n = 3.

Article Snippet: The resulting mixture was treated at 37°C for 20 min. For the analysis of NRT2 and NRT3, a second buffer was used (0.05 M Tris-HCl, pH 6.8, 4% (w/v) SDS, 12% (w/v) glycerol, 2% (v/v) 2- mercaptoethanol, and 0.05% (w/v) bromophenol blue), and samples were boiled for 5 min. Five micrograms of ECL TM Plex Fluorescent Rainbow Markers (GE Healthcare Amersham TM ) were used for each SDS-PAGE analysis.

Techniques: Western Blot, Software

Journal: Frontiers in Plant Science

Article Title: Time-Resolved Investigation of Molecular Components Involved in the Induction of NO 3 – High Affinity Transport System in Maize Roots

doi: 10.3389/fpls.2016.01657

Figure Lengend Snippet: Separation of PM H + -ATPase complexes by non-denaturing Deriphat-PAGE followed by SDS-PAGE. (A) Western blot analysis performed using antibodies anti-PM H + -ATPase. The molecular weight of each protein marker (ECL Plex Fluorescent Rainbow Markers) was reported on the right side. (B) Densitometric analysis of PM H + -ATPase. Data are expressed as ratio of corresponding 0 h sample considering the sum of all three forms (signals were quantified using PDQuest TM 2-D Analysis Software, Bio-Rad). Data are the means ± SE, n = 3.

Article Snippet: The resulting mixture was treated at 37°C for 20 min. For the analysis of NRT2 and NRT3, a second buffer was used (0.05 M Tris-HCl, pH 6.8, 4% (w/v) SDS, 12% (w/v) glycerol, 2% (v/v) 2- mercaptoethanol, and 0.05% (w/v) bromophenol blue), and samples were boiled for 5 min. Five micrograms of ECL TM Plex Fluorescent Rainbow Markers (GE Healthcare Amersham TM ) were used for each SDS-PAGE analysis.

Techniques: SDS Page, Western Blot, Molecular Weight, Marker, Software

Journal: Vaccines

Article Title: Vaccination with Ectoparasite Proteins Involved in Midgut Function and Blood Digestion Reduces Salmon Louse Infestations

doi: 10.3390/vaccines8010032

Figure Lengend Snippet: Antibody response in vaccinated fish. ( A ) The recombinant proteins were produced in E. coli and analyzed by polyacrylamide gel electrophoresis (PAGE) and Western blot (WB). Ten micrograms of recombinant proteins was loaded onto a 12% SDS-polyacrylamide gel and stained with Coomassie Brilliant Blue or transferred to a nitrocellulose membrane. For Western blot analysis, pooled sera collected from vaccinated salmons before salmon louse infestation at approximately week 10 post-initial vaccination were used as primary antibodies. The membrane was then incubated with mouse anti-rainbow trout antibodies and revealed with a goat anti-mouse IgG-HPR conjugate. ( B ) Antibody titers were determined in vaccinated and control fish by ELISA. Values of D.D. at 450 nm in vaccinated and control groups were compared by Student’s t -test with Welch’s correction for unequal variances ( p = 0.05).

Article Snippet: Plates were washed three times with PBS/0.05% Tween 20, and 100 μL/well of mouse anti-rainbow trout antibodies (Bio-Rad Laboratories, Inc.), diluted (1:1000, v / v ) in blocking solution, was added and incubated for 1 h at RT.

Techniques: Recombinant, Produced, Polyacrylamide Gel Electrophoresis, Western Blot, Staining, Membrane, Incubation, Control, Enzyme-linked Immunosorbent Assay