Puromycin Rabbit mAb (A23031)

Western blot analysis of various lysates using Puromycin Rabbit mAb (A23031) at 1:8000 dilution incubated overnight at 4℃. 293F and Raw 264.7 cells were treated by puromycin (20 μg/mL) at 37℃ for 4 hours.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 60s.

Immunohistochemistry analysis of paraffin-embedded Mouse heart tissue using Puromycin Rabbit mAb (A23031) 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.

Immunofluorescence analysis of HeLa cells (treated with puromycin) and HeLa cells (untreated) using puromycin Rabbit mAb (A23031) at dilution of 1:200 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of RAW 264.7 cells (treated with puromycin) and RAW 264.7 cells (untreated) using puromycin Rabbit mAb (A23031) at dilution of 1:200 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunoprecipitation analysis of 300ug extracts of 293T+puromycin cells using 3ug puromycin antibody (A23031). Western blot was performed from the immunoprecipitate using puromycin antibody (A23031) at a dilution of 1:5000.

Flow cytometry:1X10^6 293T cells (negative control, left) and 293T cells(treated with puromycin, right) were intracellularly-stained with puromycin Rabbit mAb(A23031, 2 μg/mL, orange line) or ABflo® 488 Rabbit IgG isotype control (A22069, 2 μg/mL, blue line). Non-fluorescently stained cells was used as blank control (red line).

Flow cytometry:1X10^6 293T cells(treated with puromycin) cells were intracellularly-stained with ABflo® 488 Rabbit IgG isotype control (A22069, 2 μg/mL, left) or puromycin Rabbit mAb(A23031, 2 μg/mL, right).

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(A23031, 2 μg/mL, orange line) or ABflo® 488 Rabbit IgG isotype control (A22069, 2 μg/mL, blue line). Non-fluorescently stained cells was used as blank control (red line).

Flow cytometry:1X10^6 Raw264.7 cells(treated with puromycin) were intracellularly-stained with ABflo® 488 Rabbit IgG isotype control (A22069, 2 μg/mL, left) or puromycin Rabbit mAb(A23031, 2 μg/mL, right).

Flow cytometry:1X10^6 C6 cells (negative control, left) and C6 cells(treated with puromycin, right) were intracellularly-stained with puromycin Rabbit mAb(A23031, 2 μg/mL, orange line) or ABflo® 488 Rabbit IgG isotype control (A22069, 2 μg/mL, blue line). Non-fluorescently stained cells was used as blank control (red line).

Flow cytometry:1X10^6 C6 cells(treated with puromycin) were intracellularly-stained with ABflo® 488 Rabbit IgG isotype control (A22069, 2 μg/mL, left) or puromycin Rabbit mAb(A23031, 2 μg/mL, right).