Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts (MEFs))
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount
25000 cells
Specification
Mouse embryonic fibroblasts (MEFs)
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 3 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
1/2000
Secondary antibody
Dilution
1/15000
Name
Non-Abcam antibody was used: IRDye 800CW Goat anti-rabbit IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Detection
Detection method
Near-Infrared Fluorescence Imaging (NIR)
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: 52, 40 kDa Non-specific: 100, 110 kDa
Additional data
Additional Notes
We used both anti-FLIP and anti-beta actin (ab6276) antibodies for the western blot. The 43 kDa red band is the beta actin band. The FLIP antibody itself detected a 52 kDa (full-length FLIP) and a 40 kDa (p43-FLIP) green bands. Two non-specific (100 &110 kDa) bands were also observed at high molecular weight region.
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
提交于 May 23 2016