Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (4-15% SDS gradient gel)
Sample
Mouse Cell lysate - whole cell (HEK293H, COS7, mouse hindbrain lysate E14)
Specification
HEK293H, COS7, mouse hindbrain lysate E14
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% Milk
Dilution
1/1000
Secondary antibody
Name
Non-Abcam antibody was used: goat anti rabbit IgG-HRP coupled
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/20000
Detection
Detection method
ECL+
Bands
Specific: 150 (GFP tagged 170) kDa Non-specific: 20, 50 kDa
Exposure
5 minute(s) and 0 second(s)
Positive control
HEK293/HeLa cells overexpressing human or mouse srGAP1,2,3, untransfected COS7 (specificity of detection doubtful), embryonic mouse hindbrain full lysate (E14)
Additional data
Additional Notes
cross reactivity with srGAP3 in HEK239H cells overexpressing human srGAP2 or srGAP3. No cross reactivity with srGAP1
Abcam response
Thank you very much for your valuable data. It appears that under these experimental conditions ab124958 cross-reacts with srGAP3. We are unable to verify the identity of the non specific band at 20.5kDa in srGAP2 overexpressing HEK293H cells, and welcome further customer feedback. Please contact scientific support for more information.
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.
DR. Alain Chedotal
Verified customer
提交于 May 09 2013