Cell Signaling Technology
Notch3 Antibody
2889
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Mammalian cells were cultured up to 70 percent confluence, serum starved for 12h and treated with growth factor. Proteins were extracted in RIPA buffer and resolved in 8% SDA-PAGE. Resolved proteins were western transferred on nitrocellulose membrane and probed to test role of treatment on NOTCH3 cleavage. Antibody selection was based on merit showed in antibody flayer.
Western Blot
Mammalian whole cell lysate
1:1000 (5% Skimmed milk in TBST), Overnight at 4 degrees Celsius
5% Skimmed milk in TBST (2h)
1:4000 (5%skimmed milk in TBST), 1h at RT
NA
ECL
This antibody gave a very bright signal of uncleaved NOTCH3 at the correct molecular weight. However, Cell Signaling's antibody could not detected cleaved NOTCH3 at 90 kDa in test mammalian cell lines.
None
Got very clear and bright signal
Good antibody