Journal: ACS chemical biology

Article Title: A Chemical Biology Solution to Problems with Studying Biologically Important but Unstable 9-O-Acetyl Sialic Acids

doi: 10.1021/acschembio.6b00928

Figure Lengend Snippet: Sialoglycan microarray binding specificity studies of human Siglec-9 (hSiglec-9-Fc, hSiglec-9), porcine torovirus hemagglutinin-esterase (PToV), and bovine coronavirus hemagglutinin-esterase (BCoV) (both PToV and BCoV were mutated to ablate their esterase activity) towards Neu5Ac9NAcα3Galβ4GlcβProNH2 (black columns), Neu5,9Ac2α3Galβ4GlcβProNH2 (gray columns), and Neu5Acα3Galβ4GlcβProNH2 (white columns) without esterase treatment (A) or treated with esterase active PToV (B).

Article Snippet: Glycan Microarray Screening Glycan microarrays were fabricated using epoxide-derivatized slides (Corning by Thermo Fisher Scientific) and Arrayit SpotBot® Extreme Microarray Spotter as previously described.

Techniques: Microarray, Binding Assay, Activity Assay

Journal: Microarrays

Article Title: Development and Optimization of a Thrombin Sandwich Aptamer Microarray

doi: 10.3390/microarrays1020095

Figure Lengend Snippet: Specificity of aptasensor for thrombin. Fluorescence signal (635 nm) recorded on thrombin aptamer microarray after the incubation of the different indicated proteins with TBA2-Cy5 (protein and TBA2 final concentration: 500 nM and 1 mM, respectively); ( a ) thrombin, blank (TBA2-Cy5 only), BSA, lysozyme and VEGF and ( b ) corresponding microarray images.

Article Snippet: TBA1 aptamers were loaded into microarray plates and submitted to the Microarray Spotter (Versarray Chip Writer Pro System, BioRad) for slide printing, using Telechem SMP3 microspotting pins (Arrayit, Sunnyvale, CA, USA).

Techniques: Fluorescence, Microarray, Incubation, Concentration Assay

Journal: Microarrays

Article Title: Development and Optimization of a Thrombin Sandwich Aptamer Microarray

doi: 10.3390/microarrays1020095

Figure Lengend Snippet: Effect of incubation time. Fluorescence signal (635 nm) recorded on TBA1 microarray ( a ) after the incubation of thrombin and TBA2-Cy5 for different times (180–120–90–60–30–15–5 min) and ( b ) corresponding microarray images.

Article Snippet: TBA1 aptamers were loaded into microarray plates and submitted to the Microarray Spotter (Versarray Chip Writer Pro System, BioRad) for slide printing, using Telechem SMP3 microspotting pins (Arrayit, Sunnyvale, CA, USA).

Techniques: Incubation, Fluorescence, Microarray

Journal: Microarrays

Article Title: Development and Optimization of a Thrombin Sandwich Aptamer Microarray

doi: 10.3390/microarrays1020095

Figure Lengend Snippet: Effect of serum. Fluorescence signal (635 nm) recorded on TBA1 microarray ( a ) after the incubation of thrombin and TBA2-Cy5 complexes in presence of different Fetal Bovine Serum (FBS) dilutions (0%–10%–20%–30%–40%–50%–65%) and ( b ) corresponding microarray images.

Article Snippet: TBA1 aptamers were loaded into microarray plates and submitted to the Microarray Spotter (Versarray Chip Writer Pro System, BioRad) for slide printing, using Telechem SMP3 microspotting pins (Arrayit, Sunnyvale, CA, USA).

Techniques: Fluorescence, Microarray, Incubation

Journal: Microarrays

Article Title: Development and Optimization of a Thrombin Sandwich Aptamer Microarray

doi: 10.3390/microarrays1020095

Figure Lengend Snippet: Direct and indirect method for thrombin detection. ( a ) Fluorescence signal (635 or 532 nm) plot of thrombin aptamer microarray after the incubation of thrombin and TBA2-Cy5 (direct method, red squares, 635 nm) or thrombin and TBA2-biotin plus Cy3-streptavidin (indirect method, green dots, 532 nm) and ( b ) relative microarray images. Tested thrombin dilutions were: 200 nM–100 nM–50 nM–10 nM–5 nM–1 nM–0 nM. TBA2-Cy5 (red squares) or TBA2-biotin (green dots) concentration was 400 nM. For the indirect method the microarray slide was incubated for 1 h with Cy3-streptavidin, for a total incubation time of two hours in all cases.

Article Snippet: TBA1 aptamers were loaded into microarray plates and submitted to the Microarray Spotter (Versarray Chip Writer Pro System, BioRad) for slide printing, using Telechem SMP3 microspotting pins (Arrayit, Sunnyvale, CA, USA).

Techniques: Fluorescence, Microarray, Incubation, Concentration Assay

Journal: Microarrays

Article Title: Development and Optimization of a Thrombin Sandwich Aptamer Microarray

doi: 10.3390/microarrays1020095

Figure Lengend Snippet: Thrombin calibration curves in FBS solution. ( a ) fluorescence signal (532 nm) plot of TBA1 microarray in 20% FBS solution after the incubation of thrombin and TBA2-biotin plus Cy3-streptavidin and ( b ) corresponding microarray images. Tested thrombin dilutions were: 50 nM–10 nM–5 nM–1 nM–0.5 nM–0 nM, with a fixed TBA-2 biotin concentration of 200 nM.

Article Snippet: TBA1 aptamers were loaded into microarray plates and submitted to the Microarray Spotter (Versarray Chip Writer Pro System, BioRad) for slide printing, using Telechem SMP3 microspotting pins (Arrayit, Sunnyvale, CA, USA).

Techniques: Fluorescence, Microarray, Incubation, Concentration Assay

Journal: BMC Genomics

Article Title: Signal transduction-related responses to phytohormones and environmental challenges in sugarcane

doi: 10.1186/1471-2164-8-71

Figure Lengend Snippet: Validation of microarray results by quantitative PCR analysis. The y axis refers to the relative expression ratio between treated samples versus the control (untreated sample). (A) and (B) Real-time PCR results for ABA and MeJA treatments, respectively. The ratios were calculated in relation to the sample from untreated plants (0h). Transcript levels of the selected genes were profiled throughout the treatment time course. Also shown are the results for plant-endophytic bacteria association (C), drought (D), phosphate starvation (E) and herbivory (F). For these treatments, the real-time PCR reactions were carried out exclusively for the experimental point(s) in which the gene was considered differentially expressed. Only validated results are shown here. The RNA samples used in the real-time PCR experiments are from a third biological sample. All reactions were carried out in parallel and each reaction was performed in triplicates. Error bars were calculated as described previously [135]. Herb. = sample from plants inoculated with Herbaspirillum seropedicae and Herbaspirillum rubrisubalbicans ; Gluc. = sample from plants inoculated with Gluconacetobacter diazotrophicus . The transcript levels for the reference genes were verified to not vary in response to the treatments. The reference genes used encode a polyubiquitin gene (SCCCST2001G02.g [CA179923]) for the ABA and drought dataset, a GAPDH for the MeJA and herbivory dataset (retrieved from [38]), a 25S rRNA for the endophytic inoculation (retrieved from [38]) and a 14-3-3 gene (SCCCLR1048F12.g [CA119519]) for phosphate starvation

Article Snippet: Microarrays were constructed by arraying cDNA fragments on DMSO optimized metal-coated glass slides (type 7, GE Healthcare) using the Generation III Microarray Spotter (Molecular Dynamics).

Techniques: Biomarker Discovery, Microarray, Real-time Polymerase Chain Reaction, Expressing, Control, Bacteria