Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Embryonic Fibroblasts (MEFs))
Loading amount
7 µg
Specification
Mouse Embryonic Fibroblasts (MEFs)
Gel Running Conditions
Non-reduced Denaturing (4-12% Bis-Tris gradient gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Other product details
Concentration
1 µg/ml
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 21°C · Diluent: Tris-Buffered Saline with 0.05% Tween 20
Secondary antibody
Name
Non-Abcam antibody was used: sheep anti-mouse IgG
Host species: Sheep
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Sheep
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/20000
Detection
Detection method
ECL+
Exposure
2 second(s)
Bands
Specific: just above 80 kDa
Positive control
Whole cell lysates from primary Mouse Embryonic Fibroblasts (MEFs) that had been immortalized by transduction with a virus harboring the gene encoding the large T antigen.
Negative control
Whole cell lysates from primary Mouse Embryonic Fibroblasts (MEFs) that had not been immortalized.
Additional data
Additional Notes
Lane 1: Whole cell lysate from WT MEFs that were virally transduced with a gene encoding the large T antigen.
Lane 2: Whole cell lysate from WT MEFs that were not transduced.
Samples were lysed in [125mM NaCl, 25mM MOPS pH 7.5, 1% tritonX-100] and spun through Qiashredder spin filter (Qiagen) to reduce viscosity due to genomic DNA. After SDS-PAGE, the samples were blotted onto PVDF membrane.
Lane 2: Whole cell lysate from WT MEFs that were not transduced.
Samples were lysed in [125mM NaCl, 25mM MOPS pH 7.5, 1% tritonX-100] and spun through Qiashredder spin filter (Qiagen) to reduce viscosity due to genomic DNA. After SDS-PAGE, the samples were blotted onto PVDF membrane.
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.
Mr. Adam Hoffhines
Verified customer
投稿 Mar 25 2008