Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Human Cell lysate - whole cell (HeLa, RPE-1)
Specification
HeLa, RPE-1
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS in 5% milk
Dilution
1/1000
Secondary antibody
Name
Non-Abcam antibody was used: LICOR, anti rabbit 800CW
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye 800CW
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye 800CW
Dilution
1/20000
Detection
Detection method
Near infrared fluorescence
Bands
Specific: 66, 55, 47 kDa
Exposure
7 minute(s) and 0 second(s)
Additional data
Additional Notes
Lysis buffer: 30mM HEPES, 175mM NaCl, 2,5mM MgCl2, 10% glycerol, 1mM PMSF, 1mM DTT, 0.5 % NP40, complete protease inhibitor (Roche)
Lane 1: 25 ug HeLa extract
Lane2: 50 ug HeLa extract
Lane 3: 25 ug RPE-1 extract
Lane 4: 50 ug RPE-1 extract
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.
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提交于 Sep 20 2013