Application
Western blot
Sample
Human Cell lysate - whole cell (RS411 (leukemic cell line))
Gel Running Conditions
Reduced Denaturing (15% acrylamide)
Loading amount
1e+006 cells
Treatment
5μM MG132
Specification
RS411 (leukemic cell line)
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C
Other product details
Incubation time
4 hour(s) and 0 minute(s) · Temperature: 20°C · Diluent: 20% blocking buffer in PBS-T
Dilution
1/1000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: Swine anti Rabbit
Host species: Rabbit
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Host species: Rabbit
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL+
Negative control
untransfected Hek293
Exposure
30 minute(s) and 0 second(s)
Bands
Specific: 17 kDa Non-specific: 10 kDa
Positive control
Hek293 cells transfected with flag-BTG2 (lane 2)
Additional data
Additional Notes
The antibody gives a clean signal but only detects overexpressed BTG2. Endogenous protein is not detected. Longer incubation times, increased concentrations or longer exposure times did not improve detection.
Lane 1: Hek293
Lane 2: Hek293 transfected with Flag-BTG2
Lane 3: RS411 cells
Lane 4: RS411 cells expressing shRNA targeting BTG2 (70%knockdown)
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 Dec 29 2010