Application
Western blot
Sample
Human Cell lysate - whole cell (MCF7 and HEK)
Loading amount
30 µg
Specification
MCF7 and HEK
Gel Running Conditions
Reduced Denaturing (8-16%)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Other product details
Dilution
1/2000
Incubation time
12 hour(s) and 0 minute(s) · Diluent: Milk
Secondary antibody
Name
Non-Abcam antibody was used: goat anti rabbit
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Dilution
1/20000
Detection
Detection method
IR
Exposure
3 minute(s) and 0 second(s)
Bands
Specific: 97 kDa Non-specific: hardly kDa
Positive control
I just loaded Cell lysates to check out the antibody, which gave a good band at the estimated size.
Additional data
Additional Notes
Lane 1: Marker - the markerband above the Hells (SMARCA6) band is 100 kDa, Hells is supposed to be 97 kDa.
Lane 2: lysate HEK cells
Lane 3: Lysate MCF7 cells
I must say that I got significantly worse results with a blot that was blocked only 1 h. Still it gives a band at the estimated size and I see a regulation of this band when transfecting my protein of interest. But there is some unspecific bands when blocked less intensively. So far I haven't tried blocking in BSA or anything else, or more milk, sorry.
Lane 2: lysate HEK cells
Lane 3: Lysate MCF7 cells
I must say that I got significantly worse results with a blot that was blocked only 1 h. Still it gives a band at the estimated size and I see a regulation of this band when transfecting my protein of interest. But there is some unspecific bands when blocked less intensively. So far I haven't tried blocking in BSA or anything else, or more milk, sorry.
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 Aug 02 2010