Application
Western blot
Sample
Human Cell lysate - whole cell (white blood cells)
Loading amount
40 µg
Specification
white blood cells
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C
Other product details
Dilution
1/1000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 2% milk, 0.1% Tween in PBS (0.01M)
Secondary antibody
Name
Non-Abcam antibody was used: Invitrogen anti-goat IgG (A21084)
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 680
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 680
Dilution
1/5000
Detection
Detection method
LI-COR Biosciences Odyssey infrared scan
Exposure
0 second(s)
Bands
Specific: 60 kDa Non-specific: 37, 50, 55, 110, 120, kDa
Negative control
Incubation in the absence of primary antibody
Additional data
Additional Notes
Image anotation:
1.Molecular ladder
2.Plasma 1
3.Plasma 2
4.White blood cells 1
5.White blood cells 2
6.HEK293 cell lysate
We use the Odyssey infra-red imaging system and an intensity 5 was used.
1.Molecular ladder
2.Plasma 1
3.Plasma 2
4.White blood cells 1
5.White blood cells 2
6.HEK293 cell lysate
We use the Odyssey infra-red imaging system and an intensity 5 was used.
Abcam response
The presence of numerous background bands with the no-primary control in lanes 2-5 suggests that the secondary antibody is binding either to some proteins in the plasma and white blood cells lysates or the blocking agent. We recommend trying blocking with 5% BSA to see if this improves the signal.
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
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提交于 Aug 23 2007