Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (5% TCA lysate preparation with glass beads accordi)
Loading amount
500000 cells
Specification
5% TCA lysate preparation with glass beads accordi
Treatment
stationary cells
Gel Running Conditions
Reduced Denaturing (15%)
Blocking step
BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Other product details
Dilution
1/250
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: Dilution 1:250 in TBS+1% BSA
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-rabbit IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000
Detection
Detection method
ECL+
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: 17 kDa
Additional data
Additional Notes
This antibody (ab18521) nicely recognizes histone H3 at an N-terminal epitope. This is different from most other available histone H3 antibodies (normally binding at C-terminal epitopes) and therefore very useful, when N-terminal detection is desired/needed.
Binding is irrespective of the acetylation status of the histone (as it recognizes histones that carry a K9,14,18R mutation, which cannot be acetylated, to comparable level as it recognizes wild type histones).
The only deficit is a quite poor detection intensity (relatively long exposure and high antibody concentration is needed, see pics for comparison with ab1791). -> therefore the rating "average"
Binding is irrespective of the acetylation status of the histone (as it recognizes histones that carry a K9,14,18R mutation, which cannot be acetylated, to comparable level as it recognizes wild type histones).
The only deficit is a quite poor detection intensity (relatively long exposure and high antibody concentration is needed, see pics for comparison with ab1791). -> therefore the rating "average"
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.
HERR DR. Tobias Eisenberg
Verified customer
投稿 Mar 29 2010