Application
Western blot
Sample
Human Cell lysate - whole cell (Lung)
Loading amount
10 µg
Specification
Lung
Treatment
10ng/mL IFN gamma for 24hrs
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Other product details
Dilution
1/5000
Incubation time
24 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 10% Milk in TBST
Secondary antibody
Name
Non-Abcam antibody was used: anti-rabbit IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
ECL
Exposure
30 minute(s) and 0 second(s)
Additional data
Abcam response
For phosphorylated proteins, we recommend blocking and diluting with a solution of BSA, 1-5%. Milk contains phospho-tyrosinses and phospho-serines; this antibody may have bound to the milk phospho-tyrosine proteins, which would account for the poor signal from JAK2 (phospho Y1007 + Y1008) on the membrane.
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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投稿 Jul 31 2007