Application
Western blot
Sample
Human Cell lysate - other (breast cancer cells (MCF7) and colon cancer cells)
Loading amount
20 µg
Specification
breast cancer cells (MCF7) and colon cancer cells
Gel Running Conditions
Non-reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/2500
Incubation time
10 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% Milk / TBST
Secondary antibody
Name
Non-Abcam antibody was used: LICOR IRDye secondary antibody
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Dilution
1/15000
Detection
Detection method
Odyssey laser scanner
Exposure
1 hour(s), 0 minute(s) and 0 second(s)
Bands
Specific: 46 kDa Non-specific: 37 kDa
Positive control
MCF7 and SW480 cell lysates. Cells were lysed using NP40 lysis buffer.
Negative control
No
Additional data
Additional Notes
Experiment was also done using 5% BSA TBST for blocking and antibody incubation but there are a lot of non-specific bands.
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投稿 Aug 03 2011