Application
Western blot
Sample
Human Cell lysate - other (Membrane fraction of Hela Cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
100000 cells
Specification
Membrane fraction of Hela Cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 20°C · Diluent: PBS + 0.05% Tween
Dilution
1/2000
Secondary antibody
Dilution
1/10000
Name
Non-Abcam antibody was used: Donkey anti-Rabbit
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Exposure
3 minute(s) and 0 second(s)
Bands
Specific: 300 kDa kDa Non-specific: 35 kDa
Additional data
Additional Notes
Tried with Milk as blocking agent and got lots of non-specific bands. BSA works better as a block but gives a high background on this membrane. Samples are from ultracentrifuged cell membrane preparations. Track 1 = 50000g pellet, Track 2 = 100000g pellet, Track 3 200000g pellet, Track 4 = Markers.
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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投稿 Sep 12 2008