Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (Vero Cells)
Loading amount
10 µg
Specification
Vero Cells
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 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: PBS Tween Milk
Secondary antibody
Name
Non-Abcam antibody was used: donkey anti rabbit
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/3000
Detection
Detection method
ECL Pierce
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: between 39 and 51 kD kDa Non-specific: None kDa
Positive control
HeLa cells
Additional data
Additional Notes
Whole cell lysates prepared from human HeLa (lanes 1 and 2) or african green monkey Vero (lanes 3 and 4) cells were prepared in 0.5% NP40 (30 minutes on ice) followed by a 14,000 rpm clarification step. 10 microgrammes of the lysates were loaded on a 4-12% NuPAGE gel and run with MOPS-SDS buffer (Invitrogen). The proteins were transfered on a nitrocellulose membrane which was incubated with the primary antibody at 1:1000 over night at 4 degrees. The blot was revealed with a classic ECL substrate (Pierce) and bands are visible after 1 second exposure. The blot shown is a 1 minute exposure. The molecular weight marker used is the SeaBlue2Plus (Invitrogen). Note: lane 2 and 4 were made from cells treated with an infectious agent that partly degrades the protein.
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提交于 Oct 24 2007