The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use a concentration of 2 µg/ml. Predicted molecular weight: 66 kDa.
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Component of the origin recognition complex (ORC) that binds origins of replication. DNA-binding is ATP-dependent, however specific DNA sequences that define origins of replication have not been identified so far. ORC is required to assemble the pre-replication complex necessary to initiate DNA replication.
Origin recognition complex, subunit 2, S. cerevisiae, homolog of antibody
Origin recognition complex, subunit 2-like (S. cerevisiae) antibody
Western blot - Anti-ORC2 antibody [SB46] (ab31930)Image from Granata M et al, PLoS Genet. 2010 Aug 5;6(8). pii: e1001047, Fig S1.
The Western blots in which the presence of Rad9 was analyzed in the total (T), soluble (S) and chromatin-enriched (Ch) fractions from wt (K699) cells, were controlled for proper fractionation of control proteins, known to remain in the soluble fraction (Tubulin) or to bind to chromatin (Orc2, ab31930).
Overlay histogram showing HeLa cells stained with ab31930 (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 (ab31930, 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.
Hiraga SI et al. Human RIF1 and protein phosphatase 1 stimulate DNA replication origin licensing but suppress origin activation. EMBO Rep18:403-419 (2017).
Read more (PubMed: 28077461) »
Granata M et al. Dynamics of Rad9 chromatin binding and checkpoint function are mediated by its dimerization and are cell cycle-regulated by CDK1 activity. PLoS Genet6: (2010).
Read more (PubMed: 20700441) »