Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T cells)
Gel Running Conditions
Reduced Denaturing (4-12% SDS-PAGE gel)
Loading amount
25 µg
Specification
HEK293T cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Other product details
Incubation time
17 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% Milk-TBST
Dilution
1/1000
Secondary antibody
Dilution
1/15000
Name
Non-Abcam antibody was used: Li-Cor anti-Rabbit
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Detection
Detection method
Odyssey Infrared Imaging System
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 25 kDa
Positive control
This sample
Additional data
Additional Notes
HEK293T cells were lysised in 1% Triton X-100 and clarified by centrifugation. The resulting lysate was separated on a 4-12% SDS-PAGE gel. Proteins were transferred on to PVDF membrane, before being blocked for 1h at room temp. Rab4 was immunoprobed for using standard procedures. The resulting immunocomplexes were detected on a Odyssey system, with scan intensity set to 7. A strong band corresponding to Rab4 was detected at approximately 25kDa.
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投稿 Jan 28 2013