The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IP: Use at 5-15µg/mg of lysate.
WB: 1/2000 - 1/10,000. Predicted molecular weight: 58 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Isoform 1: Required for ribosome biogenesis and telomere maintenance. Probable catalytic subunit of H/ACA small nucleolar ribonucleoprotein (H/ACA snoRNP) complex, which catalyzes pseudouridylation of rRNA. This involves the isomerization of uridine such that the ribose is subsequently attached to C5, instead of the normal N1. Each rRNA can contain up to 100 pseudouridine ('psi') residues, which may serve to stabilize the conformation of rRNAs. Also required for correct processing or intranuclear trafficking of TERC, the RNA component of the telomerase reverse transcriptase (TERT) holoenzyme. Isoform 3: Promotes cell to cell and cell to substratum adhesion, increases the cell proliferation rate and leads to cytokeratin hyper-expression (when overexpressed in HeLa cells).
Involvement in disease
Defects in DKC1 are a cause of dyskeratosis congenita X-linked recessive (XDKC) [MIM:305000]. XDKC is a rare, progressive bone marrow failure syndrome characterized by the triad of reticulated skin hyperpigmentation, nail dystrophy, and mucosal leukoplakia. Early mortality is often associated with bone marrow failure, infections, fatal pulmonary complications, or malignancy. Defects in DKC1 are the cause of Hoyeraal-Hreidarsson syndrome (HHS) [MIM:300240]. HHS is a multisystem disorder affecting males and is characterized by aplastic anemia, immunodeficiency, microcephaly, cerebellar hypoplasia, and growth retardation.
Belongs to the pseudouridine synthase TruB family. Contains 1 PUA domain.
Cytoplasm and Nucleus > nucleolus. Nucleus > Cajal body. Also localized to Cajal bodies.
ICC/IF image of ab93777 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 ab93777 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
Western blot - Anti-DKC1 antibody (ab93777)
All lanes : Anti-DKC1 antibody (ab93777) at 0.04 µg/ml
Lane 1 : HeLa cell lysate at 50 µg Lane 2 : HeLa cell lysate at 15 µg Lane 3 : HeLa cell lysate at 5 µg Lane 4 : 293T cell lysate at 50 µg
Detection of Human DKC1 by Immunoprecipitation in Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded)using ab93777 at 10 ug/mg lysate (LANE 1). DKC1
was also immunoprecipitated by an antibody which recognizes a downstream epitope (LANE 2). Lane 3 represents IgG Ip control. subsequent WB detection was performed using 1 ug/ml ab93777. Detection: Chemiluminescence with exposure times of 10 seconds.
All lanes : Anti-DKC1 antibody (ab93777) at 1 µg/ml
Lane 1 : HeLa cell lysate immunoprecipitated with ab 93777 at 10ug/mg lysate. Lane 2 : HeLa cell lysate immunoprecipitated with anti-DKC1 antibody at 10ug/mg lysate. Lane 3 : HeLa cell lysate immunoprecipitated with control IgG at 10ug/mg lysate.