Application
Western blot
Sample
Human Cell lysate - whole cell (Arterial endothelial cells)
Loading amount
30 µg
Specification
Arterial endothelial cells
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Other product details
Dilution
1/500
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: TBS 0,1% Tween
Secondary antibody
Name
Non-Abcam antibody was used: Donkey anti-goat
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/20000
Detection
Detection method
ECL
Exposure
30 second(s)
Bands
Specific: 37 kDa Non-specific: 35 kDa
Additional data
Additional Notes
Left lane is a control, right -- treated with GIPC1 siRNA.
Abcam response
Thank you for submitting this valuable feedback. This SiRNA experiment indicates that the antibody is specific. Howver, the image kindly provided does contain several non specific bands. We would recommend further optimization of the procedure may be required to improve results. For example, try using both the primary antibody at a lower dilution, try 1:1000 to 1:2000. It would also be important to run no primary control to assess if the background bands are from non specific binding of the secondary. The concentration of the secondary antibody may need to be reduced.
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.
Mrs. Elena Astanina
Verified customer
投稿 Apr 09 2008