The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
1/2000 - 1/10000. Detects a band of approximately 145 kDa (predicted molecular weight: 121 kDa).
Use at 2-5 µg/mg of lysate.
Regulator that plays a central role in regulation of apoptosis via its interaction with p53/TP53. Regulates TP53 by enhancing the DNA binding and transactivation function of TP53 on the promoters of proapoptotic genes in vivo.
Reduced expression in breast carcinomas expressing a wild-type TP53 protein.
Belongs to the ASPP family. Contains 2 ANK repeats. Contains 1 SH3 domain.
The ankyrin repeats and the SH3 domain are required for specific interactions with TP53.
Cytoplasm. Nucleus. Predominantly cytoplasmic. Some fraction is nuclear.
Protein phosphatase 1 regulatory subunit 13B antibody
Western blot - Anti-ASPP1 antibody (ab71163)
All lanes : Anti-ASPP1 antibody (ab71163) at 0.1 µg/ml
Lane 1 : Whole cell lysate from 293T cells at 50 µg Lane 2 : Whole cell lysate from 293T cells at 15 µg Lane 3 : Whole cell lysate from 293T cells at 5 µg Lane 4 : Whole cell lysate from Hela cells at 50 µg
Immunoprecipitation/ Western Blot of ASPP1.
Lane 1: ab71163 at 3µg/mg whole cell lysate.
Lane 2: Control IgG.
All lanes: Whole cell lysate from Hela cells at 1mg for IP, 20% of IP loaded.
Detection: Chemiluminescence with an exposure time of 10 seconds.
ICC/IF image of ab71163 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab71163, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h.Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.