The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
1/5000 - 1/15000. Detects a band of approximately 194 kDa (predicted molecular weight: 194 kDa).
Use at 2-5 µg/mg of lysate.
Use at an assay dependent dilution. PubMed: 20691152
RelevanceXrn1 is the major 5'-3' exoribonuclease involved in mRNA decay. Required for the 5'-3'processing of the G4 tetraplex-containing DNA and RNA substrates. The kinetic of hydrolysis is faster for G4 RNA tetraplex than for G4 DNA tetraplex and monomeric RNA tetraplex. Binds to RNA and DNA (By similarity). May act as a tumor suppressor protein in osteogenic sarcoma (OGS). Associates with alpha and beta tubulins (By similarity). Found in a mRNP complex with RENT1, RENT2, RENT3B and XRN1.
Whole cell lysate from HeLa (5, 15 and 50 µg for WB; 1 mg for immunoprecipitation, 20% of IP loaded) and 293T (T; 50 µg) cells. ab70259 used for western blot at 0.1 µg/ml (A) and 1 µg/ml (B) and used for IP at 3 µg/mg lysate.
XRN1 was also immunoprecipitated by ab70259. Detection was by chemiluminescence with exposure times of 30 seconds (A) and 10 seconds (B).
ICC/IF image of ab70259 stained DU145 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 (ab70259, 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.