biorobotics microgrid-ii arrayer  (BioRobotics Ltd)

Journal: Acta biomaterialia

Article Title: Single-dose combination nanovaccine induces both rapid and long-lived protection against pneumonic plague

doi: 10.1016/j.actbio.2019.10.016

Figure Lengend Snippet: Serum samples collected from C57BL/6NCrl mice (n = 12–16 per group) immunized with either saline, CDN Nanovaccine, Nanovaccine, or Combination Nanovaccine at 14, 79, and 178 DPI was analyzed for total anti-F1-V IgG antibodies against twenty-seven 14- to 17-mer linear peptides (11 amino acid overlaps) from the F1 antigen and fifty-three 15- to 17-mer linear peptides (11 or 12 amino acid overlaps) from the V antigen. The peptides were covalently bound to microarray slides as described in Materials and Methods. Each row corresponds to a specific peptide, the top row representing peptide F1 and the proceeding downward rows corresponding to each following linear peptide incrementally through peptide V53. Each column represents responses from a single mouse. The mean fluorescence intensity of serum responses to each peptide is represented by a range of color from white (no response) to purple (maximum response). The full-length F1-V fusion protein was used as a positive control, and Bacillus anthracis protective antigen (PA) and chicken egg ovalbumin (OVA) were used as negative controls. Black arrows indicate significance (p ≤ 0.05) of Combination Nanovaccine and CDN Vaccine serum compared to naïve serum at all time points evaluated. Blue arrows indicate significance (p < –0.05) of Combination Nanovaccine and CDN Vaccine serum compared to naïve serum at 179 DPI. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Peptide microarray printing and analysis Twenty-seven 14- to 17-mer linear peptides (11 amino acid overlaps) spanning the full length of F1 antigen and fifty-three 15-to 17-mer linear peptides (11 or 12 amino acid overlaps) spanning the full length of V antigen, as well as full length proteins F1-V, Bacillus anthracis protective antigen (PA), and chicken egg ovalbumin (OVA) were printed onto Nexterion Slide AL (Schott, Louisville, KY) using a BioRobotics MicroGRID II microarray printer (Genomic Solutions, Inc. Ann Arbor, MI).

Techniques: Microarray, Fluorescence, Positive Control

Journal: Nucleic Acids Research

Article Title: The Effects of Sequence Variation on Genome-wide NRF2 Binding—New Target Genes and Regulatory SNPs

doi: 10.1093/nar/gkw052

Figure Lengend Snippet: Testing the limits of NRF2 binding. Systematic multivariate ARE results by protein binding microarray for NRF2-MAFG heterodimer are shown. Tolerance for sequence variation was studied for positions 1–3 (white), 6–8 (grey) and 9–11 (dark grey) ( A ) and positions 1, 5 and 11 (seed sequences are shown in grey) ( B ). Results are depicted as measured binding relative to NQO1.ARE binding (black) (mean ± S.E.M., n = 45).

Article Snippet: The dilutions were dispensed in a 384-well plate (polypropylene plate No 267462, Nunc, N.Y, USA) and printed onto the avidin-coated glass slides with a microarray printer (BioRobotics MicroGrid II, BioRobotics Ltd, Cambridge, UK).

Techniques: Binding Assay, Protein Binding, Microarray, Sequencing

Journal: Nucleic Acids Research

Article Title: The Effects of Sequence Variation on Genome-wide NRF2 Binding—New Target Genes and Regulatory SNPs

doi: 10.1093/nar/gkw052

Figure Lengend Snippet: A SNP in FTL promoter has drastic effects of NRF2 binding and transcriptional activation. ( A ) A promoter analysis of the FTL gene at chr19 showing the location of experimentally verified NRF2 binding ARE together with dbSNP (v138) and ENCODE ChIP-seq data. ChIP-seq track displays combined MAFF and MAFK binding signals in H1-hESC (MAFK), K562 (MAFF, MAFK), HeLa-S3 (MAFK), HepG (MAFF, MAFK) and IMR90 (MAFK) cell lines. ( B ) Detailed view showing FTL ARE sequence and the SNP (rs113067944, A→C) position. ( C ) Protein binding microarray results for FTL.ARE.A and the SNP bearing FTL.ARE.C. Results are calculated as measured binding relative to NQO1.ARE binding (mean ± S.E.M, n = 39.). Scramble oligonucleotides served as negative control. ( D ) HEK-293T cells were transfected with NQO1-ARE and FTL-ARE bearing either allele A or allele C with and without NRF2 -expressing plasmids. Twenty-four h after transfection cells were treated with NRF2 inducer (L-SFN) for 16 h followed by luciferase activity measurements. An empty pGL3 promoter vector served as control and activities were normalized to β-galactosidase activity. Results are shown relative to control (mean± S.E.M, n = 4).

Article Snippet: The dilutions were dispensed in a 384-well plate (polypropylene plate No 267462, Nunc, N.Y, USA) and printed onto the avidin-coated glass slides with a microarray printer (BioRobotics MicroGrid II, BioRobotics Ltd, Cambridge, UK).

Techniques: Binding Assay, Activation Assay, ChIP-sequencing, Sequencing, Protein Binding, Microarray, Negative Control, Transfection, Expressing, Luciferase, Activity Assay, Plasmid Preparation, Control