Application
Western blot
Sample
Mouse Cell lysate - whole cell (Primary lung epithelial cells and MLE12 cells)
Loading amount
30 µg
Specification
Primary lung epithelial cells and MLE12 cells
Treatment
33nM neuregulin for 5min in primary lung epithelial cells or 0-30 min in MLE12
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Other product details
Dilution
1/500
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBS Tween 3% BSA
Secondary antibody
Name
Non-Abcam antibody was used: Anti-rabbit IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/15000
Detection
Detection method
ECL
Exposure
2 minute(s) and 0 second(s)
Bands
Specific: 180 kDa both primary kDa Non-specific: above 250kDa in MLE1 kDa
Positive control
MCF7 gave strong 180kDa band maximal at 5 min neuregulin exposure
Additional data
Additional Notes
Both MLE12 epithelial and primary fetal epithelial cells gave moderate narrow band at 180kDa. MLE12 also showed heavier broad band at about 170kDa, not seen in primary cells. Lysates probed on nitrocellulose membranes.
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Submitted Jun 18 2010