Application
Western blot
Sample
Dog Cell lysate - whole cell (MDCK)
Loading amount
15 µg
Specification
MDCK
Gel Running Conditions
Reduced Denaturing (15% tris-glycine)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Other product details
Dilution
1/500
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5 % Milk 0.2% Tween-20 in PBS
Secondary antibody
Name
Non-Abcam antibody was used: donkey anti-goat HRP
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/1000
Detection
Detection method
ECL+
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: c. 18 KDa kDa
Positive control
All lanes
Additional data
Additional Notes
Gel shows a single band at just above 16 KDa (15 % tris-glycine gel). No non-specific bands are noted. Samples are whole cell lysate (homogenised in water) from untransfected MDCK (canine kidney) cells. Samples have ben treated with an in-house synthesised small molecule that small causes a drop in the level of ARF-1 in the cell (gel shows increasing drug concentrations shown left to right and corresponding drop in ARF-1 levels).
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.
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投稿 Nov 19 2012