Application
Western blot
Sample
Mouse Cell lysate - other (Histones (acid extracted from NIH 3T3 cells))
Loading amount
50000 cells
Specification
Histones (acid extracted from NIH 3T3 cells)
Gel Running Conditions
Reduced Denaturing (AUT gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBST milk
Secondary antibody
Name
Non-Abcam antibody was used: Goat Anti-Rabbit IgG (H+L), Peroxidase Conjugated
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
ECL+
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: H3.3 kDa Non-specific: H3.1, H3.2 kDa
Additional data
Additional Notes
Histones were separated by pH (by AUT gel). H3.1, H3.2, H3.3 are indicated by arrows (for reference, please see http://www.nature.com/nprot/journal/v2/n6/full/nprot.2007.202.html). Right lane corresponds to 1 in 4 dilution of left lane. Observed many non-specific bands (including H3.1 and H3.2), as well as staining for H3.3. Two dilutions are shown.
Abcam response
Thank you for sharing this data with us. We have not previously tested this antibody on an AUT gel. This abreview shows that ab94817 detects many histone variants, under these conditions. Abcam would welcome feedback from other customers using this antibody.
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.
Abcam user community
Verified customer
投稿 Oct 31 2012