Western blot abreview for Anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) [EPR5683] antibody

Good
Abreviews
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - whole (0-4hr embryos)
Specification
0-4hr embryos
Treatment
control shRNA and SNF1A shRNA embryos
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Other product details

Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk/TBST
Dilution
1/1000

Secondary antibody

Name
Non-Abcam antibody was used: Amersham Anti-Rabbit IgG, peroxidase-linked whole
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000

Detection

Detection method
Pierce ECL
Bands
Specific: 70 kDa Non-specific: 140, 125, 38 kDa
Exposure
5 second(s)
Negative control
0-4hr old SNF1A shRNA embryos (lane 2), determined by qPCR to achieve 96% knockdown of SNF1A and exhibit phenotype
Positive control
0-4hr old EGFP shRNA embryos (lane 1)

Additional data

Additional Notes
lysis was in SDS sample buffer by homogenization with Bullet blender (Next Advance) PVDF membrane for transfer total AMPK blot below anti-pAMPK blot to monitor loading

Dr. Richelle Sopko

Verified customer

Submitted Dec 16 2013

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