Journal:

Article Title: Antibody selection for immobilizing living bacteria

doi: 10.1021/ac9014484

Figure Lengend Snippet: Immobilization of living bacteria and fabrication of bacterial cell microarray. (A) The substrate was modified with tether molecules (orange bars), followed by the covalent linking of antibody molecules (cyan Y shapes). The antibody-modified substrate was then incubated with a bacterial culture (brown ovals) to form a bacterial monolayer. The bacterial surface antigens are indicated as radial green bars. (B) The pattern of the microarray was achieved by plotting on the tether-modified substrate using a microplotter and antibody solution as “ink.” When the substrate is incubated with the bacterial culture, the bacterial cells adhere only to the antibody-modified areas and thus form a bacterial cell microarray. The beakers with orange or brown solution represent incubation with tether solution or bacterial culture, respectively.

Article Snippet: Fabrication of antibody microarray with a microplotter Antibody arrays were fabricated with a microplotter 26 (Sonoplot Inc., Madison, WI) consisting of three robotic axes which moves a micropipette tip over a large volume (∼1 m 3 ) with micron-length positional precision.

Techniques: Microarray, Modification, Incubation

Journal:

Article Title: Antibody selection for immobilizing living bacteria

doi: 10.1021/ac9014484

Figure Lengend Snippet: Reflection image of a microarray of single S. Typhimurium Δasd∷kanR H71-pHC cells prepared using DPN. Inset: an example of the MHA pattern on gold substrates before the covalent linking of antibody (lateral force image, scan size: 20 μm).

Article Snippet: Fabrication of antibody microarray with a microplotter Antibody arrays were fabricated with a microplotter 26 (Sonoplot Inc., Madison, WI) consisting of three robotic axes which moves a micropipette tip over a large volume (∼1 m 3 ) with micron-length positional precision.

Techniques: Microarray

Journal: Stem Cell Research & Therapy

Article Title: Oxidant therapy improves adipogenic differentiation of adipose-derived stem cells in human wound healing

doi: 10.1186/s13287-021-02336-3

Figure Lengend Snippet: Characterization of the macrophage activation model. a THP1 cell treatment protocol for in vitro polarization of macrophages. b Morphology (phase contrast, scale bar 100μm) of THP1-derived macrophages on day of CM harvest. c Heatmap of log2 cytokine protein levels in differentially activated macrophages. The dendrogram represents the result from a complete linkage hierarchical clustering of mean-centered expression levels based on Euclidian similarity. d mRNA-expression levels of macrophage-selective markers (M (IFNG/LPS) : CD80, IL1B, and M (IL4/IL13) : CD200R, TGM2). Data were normalized to the 18S rRNA reference gene and shown as mean –ΔCt. Asterisks indicate p values < 0.05 (*), p <0.01 (**), and p <0.001 (***). Statistical significance ( n =5) was determined by using one-way ANOVA and Dunnett’s test for multiple comparison

Article Snippet: Macrophage-CM was loaded onto RayBio® Human Cytokine G5 Antibody Microarray Glass Chip (RayBiotech, Norcross, GA) which facilitates the detection of 80 targets.

Techniques: Activation Assay, In Vitro, Derivative Assay, Expressing

Journal: Experimental Biology and Medicine

Article Title: Annona atemoya leaf extract ameliorates cognitive impairment in amyloid-β injected Alzheimer’s disease-like mouse model

doi: 10.1177/1535370219886269

Figure Lengend Snippet: Molecular mechanisms responsible for anti-AD effects of AAL extract. (a) HT22 cells were exposed to H2O2 in the absence or presence of AAL extract (50 μg/mL) for 6 h. Antibody microarray assay was performed using the phospho-specific antibody microarray slide (Full Moon BioSystems). For data acquisition, GenePix 4100A scanner (Axon Instrument, USA) was used. The normalization data were analyzed using Genowiz 4.0™ (Ocimum Biosolutions). The phosphorylation ratio was calculated and represented as fold changes of indicated phosphoproteins after H2O2 treatment normalized to total protein expression (upper panel). Total protein quantification is shown (lower panel). (b) HT22 cells were exposed to H2O2 with or without various concentrations of AAL extract (0, 12.5, 25, or 50 μg/mL) for 6 h. Cell lysates were prepared from HT22 cells and equal amounts of protein were subjected to Western blotting using anti-phospho-CaMK2 β/ν/δ, GRK2, EGFR, Myc, FER, caveolin-1, NFκB p65, and MLRN 2 antibodies to validate the Ab microarray. (c) Protein extracts were prepared from hippocampal tissues in an Aβ-induced AD mouse model. Vehicle or various concentrations of AAL extract (50, 100, or 200 mg/kg) were administered to Aβ mice for 23 days. Western blotting was performed for anti-phospho-EGFR and phospho-GRK2. The validity of the two phospho-antibodies was determined using pre-stained protein marker (Bio-Rad). GAPDH was used as an internal control. Shown blots are representative results from three independent experiments.

Article Snippet: Internal positive controls GAPDH and β-actin were constitutively expressed in all samples. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 6. caption a7 Molecular mechanisms responsible for anti-AD effects of AAL extract. (a) HT22 cells were exposed to H 2 O 2 in the absence or presence of AAL extract (50 μg/mL) for 6 h. Antibody microarray assay was performed using the phospho-specific antibody microarray slide (Full Moon BioSystems).

Techniques: Microarray, Expressing, Western Blot, Staining, Marker