NuPAGE MOPS running buffer (Invitrogen) was used, as well as pre-stain plus (Fermentas) protein ladder. Antibody production. The various antigens used in the animal immunizations are listed in Supplementary Table S1 and immunizations were carried out commercially (Agrisera, Vännäs, ...
www.nature.com/scientificreports OPEN Anti-high mobility group box-1 (HMGB1) antibody inhibits hemorrhage-induced brain injury received: 28 November 2016 accepted: 09 March 2017 Published: 10 April 2017 and improved neurological deficits in rats Dengli Wang1, Keyue Liu1, Hidenori Wake1, ...
Dead cells were gated out of the analysis by staining with the viability dye VIVID (Invitrogen). The percentage of cells labelled with each antibody was calculated in comparison with cells stained with isotype control antibody. Analysis gates for each antibody were set by using fluorescence minus ...
Plasmids for each antibody variant were transfected into Expi293 cells (Invitrogen) supernatant at a 1:3 heavy:light containing secreted ratio. After antibodies incubating at was collected b37y °cCenwtriitfhug8a%tioCnOa2t at 125 rpm for 6 days, the 500 g for 15 min at 25°C....
Following incubation, cells were pelleted and washed two times in 200 µL assay buffer before the addition of secondary antibody: rabbit polyclonal antibody to human albumin (Abcam, #ab2406). Plates were incubated and the wash routine was repeated before the tertiary antibody, goat anti-...
The separated proteins were transferred to a 0.45-μm nitrocellulose membrane (Bio-Rad, USA) and probed using Anti-MUC1-CD (AB2) rabbit polyclonal antibody (catalogue code AV41446, Sigma-Aldrich, USA) diluted 1:250 and Anti-β-actin mouse monoclonal antibody diluted 1:5000. Western blot ...
Using a T-cell transfer model of uveitis, recent study by Salomon group proposed an approach to achieve suppression of ocular inflammation by local intravitreal injection of pre-activated polyclonal Treg cells [23]. However, local delivery of retinal antigen-specific Treg cells to the inflamed eyes...
As a control, because the TSP1 antibody (rabbit polyclonal Ab8) cross-reacts with TSP1 of mouse origin, mRNA levels of mouse TSP1 was also measured, to assure that host derived TSP1 was not the source of increased TSP1 production in the treated tumors via infiltrating cells and blood (...
and then incubated with rabbit polyclonal anti-LC3B antibody (1:200 dilution) for 1 h. After extensive washing with DPBS, the cells were incubated with Alexa Fluor 488-conjugated goat anti-rabbit IgG (1:300 dilution, Invitrogen) for 1 h. The cells were counterstained with DAPI and obse...
Mifepristone (MF) has been largely used in reproductive medicine due to its capacity to modulate the progesterone receptor (PR). The study of MF has been expanded to the field of oncology; yet it remains unclear whether the expression of PR is required f