Cell Signaling
JAK1 Antibody
3344
6G4
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Browse Similar Products: JAK1 Flow Cytometry Antibodies
I generated a JAK1 knockout cell line and wanted to confirm that the JAK1 protein was completely depleted from these cells using a Western Blot. The antibody worked well and gave me a robust band in the expected size range in the positive control cell lines (the JAK1 knockout cell lines were negative as expected). However, there were some non-specific bands in the upper portion of the membrane (~180 kDa). These were not an issue as they were well outside of the expected size range, but do indicate some non-specific binding of the antibody (at least in the cancer cell lines I was using).
Western Blot
Cal27
1:1000 dilution of primary antibody in 1% BSA incubated overnight at 4 degrees celsius on a slow rocker.
5% milk
1:1000 goat-anti-rabbit HRP conjugated antibody suspended in 1% milk for one hour at room temperature
N/A
Film
As mentioned in the previous description, the antibody gave a robust band in the expected size range (~130 kDa). However, there were some non-specific bands in the upper portion of the membrane (~180 kDa) that indicate non-specific antibody binding. My positive controls had robust JAK1 expression, and my JAK1 knockout cells lost JAK1 expression as expected.
Robust bands in expected size range.
Non-specific bands in upper size range (180 kDa)
This is a good JAK1 antibody for quantifying total protein!