Western blot abreview for Anti-Bcl-2 antibody [E17]

Poor
Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
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

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.
Username

Mr. John Kelly

Verified customer

Submitted Dec 28 2007