RAP1A Antibody (1D2-1C64)

Immunocytochemistry/ Immunofluorescence: RAP1A Antibody (1D2-1C64) [NBP2-22526]

Immunocytochemistry/Immunofluorescence: RAP1A Antibody (1D2-1C64) [NBP2-22526] - Analysis of Rap1 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% BSA/TBST for 15 minutes at room temperature. Cells were probed with a Rap1 monoclonal antibody at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye.

Immunoprecipitation: RAP1A Antibody (1D2-1C64) [NBP2-22526]

Immunoprecipitation: RAP1A Antibody (1D2-1C64) [NBP2-22526] - Analysis of Rap1 was performed on NIH3T3 cells. Antigen-antibody complexes were formed by incubating 750ug of NIH3T3 cell lysate with 2ug of a Rap1 monoclonal antibody overnight on a rocking platform at 4C. The immune complexes were captured on 50ul Protein A/G Plus Agarose, washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer. Eluted sample (right lane) and 25ug of NIH3T3 control lysate (left lane) were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hour. The membrane was probed with a Rap1 monoclonal antibody at a dilution of 1:500 overnight rotating at 4C, washed in TBST, and probed with Clean-blot IP Detection Reagent at a dilution of 1:2500 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura.