Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293 cells)
Loading amount
1 µg
Specification
HEK293 cells
Gel Running Conditions
Non-reduced Denaturing (10% gel)
Blocking step
BSA as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Other product details
Dilution
1/500
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: 1 X TBS + 0.1% Tween
Secondary antibody
Name
Non-Abcam antibody was used: Anti-mouse
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000
Detection
Detection method
ECL+
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 45 kDa Non-specific: 42 kDa
Positive control
N/A
Negative control
Bacterial sample
Additional data
Additional Notes
WCE from HEK293 cells, cell pellet of approx 0.1g was lysed and approx 20% loaded. The blot was stripped once and reprobed with anti-Nap1l1 to test it. On overloaded lanes there is an extra band running ~5kDa below the band of expected size. It is clearly of a lower intensity. The bands came up quickly, so I expect a more dilute Ab mix would work, and perhaps reduce the background band.
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.
Abcam user community
Verified customer
投稿 Jul 28 2010