Cy2 goat anti mouse igg

Cells were cultured on collagen‐coated glass coverslips for 24 hour and then rinsed with PBS twice before fixation with 4% formaldehyde for 20 minute at 37°C. Subsequently, cells were rinsed with PBS for three times and permeabilized with 0.2% Triton X‐100 for 10 minute. The cells were incubated with PBS containing 1% BSA for 30 minute and then incubated with the primary antibody at 4°C overnight. Afterwards, cells were washed and incubated with fluorophore‐conjugated secondary antibodies (Cy3™‐Goat Anti‐Rabbit IgG or Cy2™‐Goat Anti‐Mouse IgG Jackson Immunoresearch) for 2 hour and then stained with DAPI for 5 minute. After the final wash, a fluorescent microscope was used (Nikon, Japan) to collect images.

C1r mouse monoclonal antibody (mAb) (#sc-514105), goat anti-rabbit IgG-HRP (#sc-2004) and goat anti-mouse IgG-HRP (#sc-2005) were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). CD11b/c mouse mAb (ab1211) was purchased from Abcam (Waltham, MA, USA). CD11b rabbit polyclonal antibody (pAb) (#NB110–89474) and GFAP rabbit pAb (#NB300–141) were procured from Novus Biologicals (Centennial, CO, USA). GAPDH rabbit mAb (#2118S) was purchased from Cell Signaling Technology (Danvers, MA, USA). PathoGreen Histofluorescent stain (#80027–5) was purchased from Biotium (Fremont, CA, USA). Cy3-goat anti-rabbit IgG (#111–165-144) and Cy2-goat anti-mouse IgG (#115–225-146) were procured from Jackson ImmunoResearch Laboratories (West Grove, PA, USA).