WB
Western blot analysis of lysates from RAW264.7 cells, using Puromycin Rabbit mAb at 1:8000 dilution. Raw264. 7 cells were treated by puromycin (20ug/ml) at 37℃ for 4 hours.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.
Lysates/proteins: 25ug per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit.
Exposure time: 180s.
IHC
Immunohistochemistry analysis of paraffin-embedded Mouse heart tissue using Puromycin Rabbit mAb at a dilution of 1:400 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.
ICC/IF
Immunofluorescence analysis of HeLa cells (treated with puromycin) and HeLa cells (untreated) using puromycin Rabbit mAb at dilution of 1:200 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) at 1:500 dilution. Blue: DAPI for nuclear staining.
FC
Flow cytometry:1X10^6 Raw264.7 cells (negative control,left) and Raw264.7 cells(treated with puromycin,right) were intracellularly-stained with puromycin Rabbit mAb(2ug/mL,orange line) or ABflo® 488 Rabbit IgG isotype control (2ug/mL,blue line). Non-fluorescently stained cells was used as blank control (red line).