Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Other product details
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk
Dilution
1/1000
Secondary antibody
Dilution
1/20000
Name
Non-Abcam antibody was used: Jackson anti-mouse heavy and light
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
SuperSignal West Pico
Negative control
Lane 1: Untransfected HEK293 cells.
Exposure
30 second(s)
Bands
Specific: 65 kDa Non-specific: 80, 90, 50 kDa
Positive control
Lane 2: HEK293 Cells transfected with an HA- tagged protein that runs at approximately 65kDa
Additional data
Additional Notes
Strong non-specific bands in this application between 75 and 100kDa would prevent use of this antibody for HA-tagged proteins in that size range.
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 Mar 03 2015