Journal: Scientific Reports

Article Title: Phosphate-regulated expression of the SARS-CoV-2 receptor-binding domain in the diatom Phaeodactylum tricornutum for pandemic diagnostics

doi: 10.1038/s41598-022-11053-7

Figure Lengend Snippet: Phosphate-regulated expression of algae codon-optimized RBD from the HASP1 promoter. ( a ) RBD expression in response to varying phosphate concentrations measured over time. Shown is a coomassie-stained gel (top) of whole cell extracts from cultures harbouring pSS2 at day 0, 3 and 5 post inoculation into media with 0%, 1%, 10% or 100% of phosphate (Supplementary Table ). RBD expression was assessed by western blotting (bottom) using a polyclonal anti-RBD antibody. The positive control (+) is 5 ng of commercially available RBD purified from HEK293 cells. M, BLUelf Prestained Protein Ladder. ( b ) RBD expression in a 5 L bioreactor. The top image is a coomassie-stained gel of whole cell lysates sampled at the indicated day, while the bottom image is a western blot using a polyclonal anti-RBD antibody. M, BLUelf Prestained Protein Ladder. ( c ) Plot of phosphate (filled circles) and iron (filled diamonds) as a function of growth over (open circles, solid line) and cell number (open circles, dashed line) over time in a photobioreactor. Phosphate and iron are plotted as parts per million (ppm), growth is plotted as absorbance readings at A670 nM, and cell number as cell count per mL of culture. Uncropped gel images for panels ( a ) and ( b ) are shown in Supplementary Figures – .

Article Snippet: Nitrocellulose membrane (Millipore) was striped at a test line location with an anti-human IgG antibody (BiosPacific Inc.) and at a control line location with a Goat anti-Rabbit polyclonal antibody (Cedarlane Inc.).

Techniques: Expressing, Algae, Staining, Western Blot, Positive Control, Purification, Cell Counting

Journal: Scientific Reports

Article Title: Phosphate-regulated expression of the SARS-CoV-2 receptor-binding domain in the diatom Phaeodactylum tricornutum for pandemic diagnostics

doi: 10.1038/s41598-022-11053-7

Figure Lengend Snippet: Purification and activity of the algae-RBD. ( a ) Representative gel images of individual chromatographic steps in the purification, starting with the HisPrep FF column on the left. For each step, numbers above the gel indicate elution fractions. M, BLUelf Prestained Protein Ladder. ( b ) Western blots with an anti-RBD or anti-6XHis antibody of the load (L), flow through (FT) and elution fraction 6 (E) from the HisPrep FF column purification of the algae-RBD construct with a TEV protease site. ( c ) Western blots as in panel ( b ) of a purification of an algae-RBD construct lacking the TEV protease sie. ( d ) Protein identification of the algae-RBD by MALDI MS. The amino acid sequence of the algae-RBD is shown and peptides identified are highlighted in yellow. ( e ) Endoglycosidase digestion assays to detect N -linked glycosylation. Purified algae-RBD or HEK293-RBD were incubated with (+) or without (−) the indicated glycosylase and visualized by western blotting with a polyclonal anti-RBD antibody. ( f ) ACE2 inhibition binding assay. Shown is a plot of absorbance readings of ACE2-bound Fc-tagged RBD at various concentrations of algae-RBD or commercially available RBD made in HEK293 cells (HEK293-RBD). Data are three independent experimental replicates with error bars representing standard deviation of the mean. Uncropped gel images for panels ( a ), ( b ), ( c ), and ( e ) are shown in Supplementary Figures – .

Article Snippet: Nitrocellulose membrane (Millipore) was striped at a test line location with an anti-human IgG antibody (BiosPacific Inc.) and at a control line location with a Goat anti-Rabbit polyclonal antibody (Cedarlane Inc.).

Techniques: Purification, Activity Assay, Algae, Western Blot, Construct, Sequencing, Glycoproteomics, Incubation, Inhibition, Binding Assay, Standard Deviation

Journal: Cell reports

Article Title: Akt3 activation by R-Ras in an endothelial cell enforces quiescence and barrier stability of neighboring endothelial cells via Jagged1

doi: 10.1016/j.celrep.2024.113837

Figure Lengend Snippet:

Article Snippet: Goat polyclonal anti-rabbit IgG HRP-conjugated , Promega , W401B.

Techniques: Virus, Control, Expressing, Plasmid Preparation, Recombinant, Isolation, MTT Assay, Western Blot, Software