Application
Western blot
Sample
Mouse Cell lysate - whole cell (B16 Melanoma)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
B16 Melanoma
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
1 hour(s) and 30 minute(s) · Temperature: 25°C · Diluent: 5% Milk
Dilution
1/500
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: Sigma Goat anti-rabbit HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
Supersignal West Pico
Exposure
3 minute(s) and 0 second(s)
Bands
Specific: none kDa Non-specific: ~70 kDa
Additional data
Additional Notes
PVDF Membrane Used
Total Cell Lysate prepared as described in product documentation, using COmplete Protease Inhibitor cocktail (Roche).
Abcam response
I'm sorry you are experiencing problems with this antibody. You may want to increase primary antibody incubation time to overnight at 4C. Only the initial block and secondary incubation should be performed in 5% nonfat dry milk; if diluting the primary antibody with a blocking reagent, use 5% BSA.
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.
Mr. John Kelly
Verified customer
投稿 Dec 28 2007