Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS cells)
Loading amount
50 µg
Specification
U2OS cells
Gel Running Conditions
Reduced Denaturing (4-20% tris-glycine)
Blocking step
normal horse serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Dilution
1/1000
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 20°C · Diluent: 5% normal horse serum in PBS
Secondary antibody
Name
Non-Abcam antibody was used: Donkey-anti-goat-HRP
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000
Detection
Detection method
ECL
Exposure
2 minute(s) and 0 second(s)
Bands
Specific: 140 kDa Non-specific: faint 100kd, 200kd kDa
Positive control
siRNA against Upf1
Negative control
control siRNA
Additional data
Additional Notes
human derived U2OS cells were treated siRNA targeting the following: Lane 1, control Lane 2, Upf1 Lane 3, GAPDH. After 4 days, cells were lysed and processed for Western blotting with the antibodies as indicated. GAPDH antibody is Abcam 9483 (see product page for Abreview). Treatment with Upf1 siRNA decreased the Upf1 Western blot signal as expected.
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.
DR. Natalie Farny
Verified customer
提交于 Oct 23 2009