Application
Western blot
Loading amount
90 µg
Gel Running Conditions
Reduced Denaturing (6% gel)
Sample
Human Cell lysate - whole cell (Human osteosarcoma U2OS cells)
Specification
Human osteosarcoma U2OS cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1% BSA in TBS-T
Dilution
1/100
Secondary antibody
Name
Non-Abcam antibody was used: Immunopure Pierce anti-mouse
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
Bands
Specific: Not sure if any kDa Non-specific: 150, 180, 200, 250 kDa
Exposure
20 minute(s) and 0 second(s)
Positive control
Whole cell extracts from U2OS cells
Additional data
Additional Notes
The image shows the entire upper part of the gel. It was cut to remove the part of the membrane that was not in contact with the gel. Any of the weak bands above the 250 kDa marker could be MLL, but it is very difficult do distinguish from the background (and the antibody was used quite concentrated 1/100 and the membrane exposed for a long time 20 min)
Abcam response
Thank you for your feedback. Ab32400 has only previously been tested when KMT2/MLL was overexpressed. These data suggest that it is unable to detect endogenous protein under these experimental conditions. For further information, please contact scientific support.
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 May 29 2013