Cell Signaling Technology
Phospho-SAPK/JNK (Thr183/Tyr185) Antibody
9251
Polyclonal
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Rat cell extract was prepared, centrifuged and the supernatant was collected. Protein was estimated in the supernatant. 40 microgram of the protein was resolved in the SDS-PAGE. The protein bands were then transferred to the nitrocellulose membrane. Nitrocellulose membrane was blocked at then incubated overnight with primary antibody (JNK1/2/3). Membrane was washed and then incubated with secondary antibody conjugated with HRP. The ECL reagent is used for producing the signal which is captured by x ray film.
Western Blot
Mammalian cell
12 h incubation at 4°C (1:1000 dilution in 5% BSA prepared in TBST)
5% blotting grade blocker (skimmed milk) in TBST
1 h at room temperature (1:4000 dilution in 5% BSA prepared in TBST)
None
ECL reagent
Specific bands were observed with the pJNK1/2/3 antibody. Some of the samples have expressed high level of pJNK1/2/3 as compared to control
N/A
The antibodies were specific and gave sharp bands.
Excellent phosphoJNK1/2/3 antibody from Cell Signaling Technology