Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (whole cell extract (done by TCA 10% protocol))
Loading amount
15 µg
Specification
whole cell extract (done by TCA 10% protocol)
Treatment
cells were grown in galactose 2% to induce HO nuclease
Gel Running Conditions
Reduced Denaturing (gel 10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/2000
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: 0.01% tween 20 in 1XPBS
Secondary antibody
Name
Non-Abcam antibody was used: ImmunoPure Antibody-Goat anti mouse
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/25000
Detection
Detection method
ECL
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 90 kDa
Positive control
none
Negative control
none
Additional data
Additional Notes
The antibody detects very well all the Rad53 phosphorylated forms in response to one irreparable double strand DNA break induced by HO nuclease overexpression. It also detects the CDK1-dependent phosphorylation induced by treatment with nocodazole that blocks cells in G2/M phase.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
DR. Achille Pellicioli
Verified customer
提交于 Nov 09 2012