The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500 - 1/1000. Detects a band of approximately 41 kDa (predicted molecular weight: 41 kDa).
1/50 - 1/100.
Use a concentration of 10 µg/ml.
The ZDHHC9-GOLGA7 complex is a palmitoyltransferase specific for HRAS and NRAS.
Highly expressed in kidney, skeletal muscle, brain, lung and liver. Absent in thymus, spleen and leukocytes.
Defects in ZDHHC9 are the cause of mental retardation syndromic X-linked ZDHHC9-related (MRXSZ) [MIM:300799]. A disorder characterized by significantly sub-average general intellectual functioning associated with impairments in adaptative behavior and manifested during the developmental period. Some patients have marfanoid habitus as an additional feature.
Belongs to the DHHC palmitoyltransferase family. ERF2/ZDHHC9 subfamily. Contains 1 DHHC-type zinc finger.
The DHHC domain is required for palmitoyltransferase activity.
ICC/IF image of ab74504 stained HeLa cells. The cells were 100% methanol fixed (5 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 ab74504 at 10µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Western blot - ZDHHC9 antibody (ab74504)
All lanes : Anti-ZDHHC9 antibody (ab74504) at 1/500 dilution
Lane 1 : extracts from COLO cells Lane 2 : extracts from COLO cells with immunising peptide at 10 µg
Lysates/proteins at 30 µg per lane.
Predicted band size : 41 kDa Observed band size : 41 kDa Additional bands at : 26 kDa,34 kDa,48 kDa. We are unsure as to the identity of these extra bands.