Application
Western blot
Sample
Human Recombinant protein (E. coli-purified GST)
Loading amount
0.05 µg
Specification
E. coli-purified GST
Gel Running Conditions
Reduced Denaturing (10% Acrylamide)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/5000
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: PBS-Tween 20 0.1%, milk 5%
Secondary antibody
Name
Non-Abcam antibody was used: ECL Anti-Rabbit IgG
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/2500
Detection
Detection method
ECL+
Exposure
5 second(s)
Bands
Specific: 25 kDa
Positive control
GST alone (25 kDa) and human METTL21D-GST (46 kDa)
Additional data
PubMed ID
23349634: View
Additional Notes
This is an In Vitro GST pulldown experiment. 100 ng of 6x-his tagged and 500 ng of GST-tagged proteins were incubated together and then purified by Glutathione beads. Samples were resolved by SDS-PAGE and western blotting shows efficiency of purification (GST blot) and evidence of direct interaction between the two protein (6xHis blot).
All proteins are of human origin but were expressed and purified from E. coli.
All proteins are of human origin but were expressed and purified from E. coli.
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投稿 Feb 21 2013