Acts as guanine nucleotide exchange factor (GEF) for RhoA GTPase. May be involved in activation of the SAPK/JNK pathway Stimulates genotoxic stress-induced RHOB activity in breast cancer cells leading to their cell death.
ab113202 staining NET1 in PC-12 cells treated with milnacipran hydrochloride (ab120755), by ICC/IF. Decrease of NET1 expression correlates with increased concentration of milnacipran hydrochloride, as described in literature. The NGF treated cells were incubated at 37°C for 6 hour in media containing different concentrations of ab120755 (milnacipran hydrochloride ) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab113202 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Western blot - NET1 antibody (ab113202)
All lanes : Anti-NET1 antibody (ab113202) at 0.4 µg/ml
Lane 1 : Jurkat Cell lysate at 50 µg Lane 2 : Jurkat Cell lysate at 15 µg Lane 3 : Jurkat Cell lysate at 5 µg
Predicted band size: 62 kDa
Exposure time: 3 minutes
Immunoprecipitation - NET1 antibody (ab113202)
Immunoprecipitation of NET1 (6 µg of antibody per mg of Jurkat cell lysate; 20% of the IP loaded). Lane 1- Immunoprecipitation with an anti-Net1 antibody that recognizes an upstream epitope; Lane 2- Immunoprecipitation with ab113202; Lane 3- Immunoprecipitation with IgG control.
Anti-Net 1 antibody (ab113202) used at 1 µg/ml for western blot. Detection: Chemiluminescence with exposure time of 3 min.