Application
Western blot
Sample
Human Cell lysate - whole cell (macrophages)
Loading amount
25000 cells
Specification
macrophages
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Dilution
1/600
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBST + 2% milk
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-rabbit (Pierce) HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/300
Detection
Detection method
ECL
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: None kDa Non-specific: 120, 80, 50, 45, 42, etc kDa
Additional data
Abcam response
Thank you for your feedback. Please let us know if the following suggestions improve your results: 1) increase blocking i.e. 2 hrs in 5% milk + 5% BSA, 2) use TRIF antibody at 1:1000 in blocking solution, 3) use secondary antibody at a higher dilution e.g. 1:1000 and 4) reduce exposure time.
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 Jan 29 2008