Application
Western blot
Sample
Mouse Purified protein (Hepa1-6 hepatoma cells)
Loading amount
1000 µg
Specification
Hepa1-6 hepatoma cells
Treatment
1 nM insulin for 15 min/no insulin
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Other product details
Dilution
1/100
Incubation time
3 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: 3% BSA
Secondary antibody
Name
Non-Abcam antibody was used: HRP anti-rabbit
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
ECL+
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: N/A kDa Non-specific: none kDa
Positive control
Immunoprecipitated novel phosphoprotein in presence of phosphorylation stimulus (insulin)
Negative control
Immunoprecipitated novel phosphoprotein in absence of phosphorylation stimulus (insulin)
Additional data
Additional Notes
Image: Protein of interest was immunoprecipitated using antibody directed at the native, non-phosphorylated protein. WB using P-Thr antibody. Lanes: 1) No insulin, 2) 1 nM insulin (phosphorylation stimulus)
1000 ug is the amount of protein used for each immunoprecipitation
1000 ug is the amount of protein used for each immunoprecipitation
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投稿 May 12 2012