Application
Western blot
Sample
Human Cell lysate - whole cell (Vascular Smooth Muscle Cells)
Loading amount
20 µg
Specification
Vascular Smooth Muscle Cells
Gel Running Conditions
Reduced Denaturing (10% Bris-Tris with MES running buffer)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Dilution
1/2000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% Milk
Secondary antibody
Name
Non-Abcam antibody was used: Donkey anti-rabbit IgG
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000
Detection
Detection method
Dura
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: 12 kDa
Additional data
Additional Notes
As the sample used was a cell lysate, the Ab is presumably detecting the propeptide form of IGF1 (12kDa) rather than the active secreted form of the protein (7kDa). The blot depicts three different samples loading evenly based on BCA assay and even loading was subsequently confirmed by a GAPDH loading control.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
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提交于 Aug 22 2008