Western blot abreview for Anti-GFP antibody [LGB-1]

Excellent
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (Hela, Huh7 and 293 cells)
Loading amount
10 µg
Specification
Hela, Huh7 and 293 cells
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris Gel)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Other product details

Concentration
0.2 µg/ml
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 24°C

Secondary antibody

Name
Non-Abcam antibody was used: ECL Mouse IgG, HRP-Linked Whole Ab
Host species: Sheep
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000

Detection

Detection method
ECL+
Exposure
10 second(s)
Bands
Specific: 27 kDa
Positive control
CMV-GFP plasmid was used as a transfection control and the GFP expression was confirmed using fluorescent microscope. For Western GFP was used to normalize the expression of another protein of interest. Thus all samples were GFP+ and were controls for themselves
Negative control
untransduced cells were used as negative control.

Additional data

Additional Notes
I used the antibody at lower concentration than recommended by the Abcam. They suggest 0.5ug/ml and I used 0.2ug/ml. It worked very well at that concentration.

Dr. Leszek Lisowski

Verified customer

Submitted Jan 06 2009

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