The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 90 kDa.
Use a concentration of 3 µg/ml.
Use a concentration of 5 µg/ml.
Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
FunctionPutative E3 ubiquitin-protein ligase. May participate in methylation-dependent transcriptional regulation. Binds to inverted 5'-CCAAT-3' box 2 in the TOP2A promoter, and activates TOP2A expression. Important for G1/S transition. May be involved in DNA repair and chromosomal stability.
Tissue specificityExpressed in thymus, bone marrow, testis, lung and heart. Overexpressed in breast cancer.
PathwayProtein modification; protein ubiquitination.
Developmental stageExpressed in fetal thymus, liver and kidney.
DomainThe RING finger is required for ubiquitin ligase activity. The YDG domain mediates the interaction with histone H3.
Post-translational modificationsPhosphorylated on serine residues. Phosphorylation may enhance DNA-binding activity. Ubiquitinated; which leads to proteasomal degradation. Polyubiquitination may be stimulated by DNA damage.
ICC/IF image of ab57083 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 (ab57083, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.
Flow Cytometry-Anti-UHRF1 antibody(ab57083)
Overlay histogram showing MCF7 cells stained with ab57083 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab57083, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
References for Anti-UHRF1 antibody (ab57083)
This product has been referenced in:
Wang F et al. UHRF1 Promotes Cell Growth and Metastasis Through Repression of p16(ink4a) in Colorectal Cancer. Ann Surg Oncol : (2012).
Read more (PubMed: 22219067) »
Rothbart SB et al. Association of UHRF1 with methylated H3K9 directs the maintenance of DNA methylation. Nat Struct Mol Biol19:1155-60 (2012).
Read more (PubMed: 23022729) »