Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (wildtype, histone H3 mutant, dot1 mutant)
Gel Running Conditions
Reduced Denaturing (BioRad AnyKD)
Loading amount
1e+006 cells
Treatment
UV exposed (200 J/m2)
Specification
wildtype, histone H3 mutant, dot1 mutant
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBS-Tween (0.05%)
Dilution
1/50
Secondary antibody
Dilution
1/3000
Name
Non-Abcam antibody was used: Goat Anti-Rabbit IgG Antibody, HRP-conjugate
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL+
Negative control
dot1 strain (no H3K79 methylation)
Exposure
20 minute(s) and 0 second(s)
Bands
Specific: 17 kDa
Positive control
WT and histone H3 mutant strain samples (mutant previously shown to have elevated H3K79 mono-methylation levels) unexposed or exposed to UV, plus additional incubation period, as indicated on figure.
Additional data
Additional Notes
No bands detectable with antibody solutions more dilute than 1:50.
Abcam response
This antibody has not yet been validated in S. cerevisiae by our lab. This researcher used our AbTrial program to test their samples risk free. If you would like to test an untested species or application please contact our scientific support team for more information about the AbTrial program.
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. Jeff Thompson
Verified customer
投稿 Jul 23 2015