Application
Western blot
Sample
Sheep Tissue lysate - whole (Liver)
Loading amount
3 µg
Specification
Liver
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/2000
Incubation time
14 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 0.05% TBST
Secondary antibody
Name
Non-Abcam antibody was used: Donkey anti-goat secondary antibody
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/3000
Detection
Detection method
ECL
Exposure
30 second(s)
Bands
Specific: 31 kDa
Additional data
Additional Notes
12 lanes in the blot are all liver samples from sheep, but there are two treatment groups. Half of the lanes were loaded with liver tissue lysate from offspring born to obese ewes, and the other half were born to Control group ewes. This antibody did show another weak band at about 50 KD, but after washing in TBST for several times (3~4) each 15 minutes, I was able to get rid of the other band.
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 Sep 21 2011