Application
Western blot
Sample
Human Cell lysate - whole cell (lung cancer cells)
Loading amount
10 µg
Specification
lung cancer cells
Treatment
with DMSO or 100 nM bortezomib for 24 hours
Gel Running Conditions
Reduced Denaturing (4-20% Bis-Tris)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Dilution
1/1000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBS + 0.1% Tween-20 + 5% BSA
Secondary antibody
Name
Non-Abcam antibody was used: goat anti-rabbit HRP
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
1 minute(s) and 0 second(s)
Bands
Specific: 25, 50, smear above kDa
Positive control
H460 cells were treated for 24 hours with DMSO or 100 nM bortezomib. Lysates were then prepared for Western analysis.
Additional data
Additional Notes
H460 cells were treated for 24 hours with DMSO or 100 nM bortezomib. Lysates were prepared directly in sample buffer (250 ul 4X NuPage sample buffer, 100 ul beta-mercaptoethanol, 650 ul H2O per ml), sonicated, boiled and loaded for Western analysis.
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.
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投稿 Jan 24 2013