Application
Western blot
Sample
Human Cell lysate - nuclear (U2OS)
Loading amount
5 µg
Specification
U2OS
Treatment
+/- 60 J/m^2 UV irradiation (harvest 15 hrs later)
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 22°C
Other product details
Dilution
1/2000
Incubation time
14 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1:1 PBS/Licor Blocking Buffer
Secondary antibody
Name
Non-Abcam antibody was used: IRDye 800CW Goat anti-Mouse IgG (H + L)
Host species: Goat
Clonality: Monoclonal
Conjugation: IRDye® 800CW
Host species: Goat
Clonality: Monoclonal
Conjugation: IRDye® 800CW
Dilution
1/10000
Detection
Detection method
Odyssey Infrared Imager
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 38 kDa
Positive control
U2OS Nuclear Lysate (Lanes 4 and 5)
Negative control
U2OS Cytosolic Lysate (Lanes 2 and 3)
Additional data
Additional Notes
Lane 1 MW Ladder in kDa (from top: 118, 90, 50, 34, 26, 19) Lane 2 and 3 Cytosolic Fractions from U2OS Cells without and with UV treatment repectively. Shows detection of CCT epsilon at 60 kDa. Lanes 4 and 5 Nuclear lysates from U2OS Cells without and with UV treatment repectively. Shows TBP at ~40 kDa and CCT epsilon at 60 kDa. Lanes 6 and 7 Insoluble nuclear fractions from U2OS Cells without and with UV treatment repectively. Shows TBP at ~ 40 kDa. Some very minor nonspecific bands are seen. This blot was blotted for both CCT epsilon and TBP.
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. Chris Tracy
Verified customer
投稿 Jan 12 2012