Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (Vero Cells)
Loading amount
10 µg
Specification
Vero Cells
Treatment
Infected with HSV KOS
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Dilution
1/1000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS Tween Milk
Secondary antibody
Name
Non-Abcam antibody was used: Anti Mouse (Amersham)
Host species: Sheep
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Sheep
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/3000
Detection
Detection method
ECL Pierce
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: between 97 and 191 kDa
Negative control
Non infected cells
Additional data
Additional Notes
Cells were infected for 16 hours with HSV KOS (wt virus), then washed with PBS and lysed with 0.5% NP40 in X buffer on ice for 15-30 minutes. The lysate was cleared by centrifugation at 14,000 rpm in a cold eppendorf centrifuged and 10 microgrammes of the lysate was loaded on a 4-12% NuPAGE gel which was run with MOPS-SDS buffer (Invitrogen) in reducing conditions. The nitrocellulose membrane was cut just under the 97 marker.
Lane 1 and 2 are uninfected and KOS infected HeLa human cells, lanes 3 and 4 are uninfected and KOS infected Vero cells.
Lane 1 and 2 are uninfected and KOS infected HeLa human cells, lanes 3 and 4 are uninfected and KOS infected Vero cells.
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 Aug 14 2007