Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS cells)
Loading amount
50 µg
Specification
U2OS cells
Gel Running Conditions
Reduced Denaturing (6-15%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Dilution
1/500
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBST with 1% BSA
Secondary antibody
Name
Non-Abcam antibody was used: Rabbit anti- goat IgG HRP
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/2000
Detection
Detection method
ECL
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 25 kDa Non-specific: 37, 90 kDa
Additional data
Additional Notes
A major band of the expected size is detected in U2OS whole cell extracts. Fractionation in cytoplasm (C), nucleoplasm (N) or chromatin pellet (P), shows that GRB2 is mainly in the cytoplasm and can be used as a marker.
Abcam response
Using milk as a blocking agent might help reduce background in whole cell lysates.
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
投稿 May 18 2011