Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts (MEFs))
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gradient gel)
Loading amount
50000 cells
Specification
Mouse embryonic fibroblasts (MEFs)
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: Li-Cor Odyssey Blocking Buffer (TBS)
Dilution
1/1000
Secondary antibody
Dilution
1/15000
Name
Non-Abcam antibody was used: IRDye 680RD Goat anti-Rabbit IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 680
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 680
Detection
Detection method
Near-Infrared Fluorescence Imaging
Exposure
6 minute(s) and 0 second(s)
Bands
Specific: 240 kDa Non-specific: 60, 75, 105 kDa
Additional data
Additional Notes
We used both anti-RNAP II pSer2 and anti-beta actin (ab6276) antibodies for the western blot. The 42 kDa green band is the beta actin band. The RNAP II pSer2 antibody itself detected a 240 kDa red band from the cell lysates.
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提交于 Feb 19 2016