Application
Western blot
Sample
Mouse Cell lysate - nuclear (Mouse embryonic fibroblasts)
Loading amount
30 µg
Specification
Mouse embryonic fibroblasts
Gel Running Conditions
Reduced Denaturing (4-16% Tris-Gly gel)
Blocking step
LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
Other product details
Dilution
1/1000
Incubation time
14 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: Odyssey® Blocker 1:1 in PBS + 0.1% Tween
Secondary antibody
Name
Non-Abcam antibody was used: AlexaFluor 680 goat anti-rabbit
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 680
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 680
Dilution
1/10000
Detection
Detection method
Fluorescent using Odyssey® scanner
Exposure
2 minute(s) and 0 second(s)
Bands
Specific: 14 kDa Non-specific: 22, 26, 32, 40-47 kDa
Positive control
Nuclear fraction
Negative control
Cytoplasmic fraction
Additional data
Additional Notes
Picture legend.
Nucl - nuclear fraction
Cyt - cytoplasmic fraction
The antibody work really great if a membrane is cut at the 20kDa mark so that no non-specific bands are detected and the background is significantly lower (data not shown)
Nucl - nuclear fraction
Cyt - cytoplasmic fraction
The antibody work really great if a membrane is cut at the 20kDa mark so that no non-specific bands are detected and the background is significantly lower (data not shown)
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 Feb 23 2012