Phospho-Akt, Cell Signaling Technology, 9271

WB dilution 1:2000

IF rating: 5 stars
IHC rating: 4 stars

For more information see Sutton et al. Cell Reports 2019, “DIRAS3 (ARHI) Blocks RAS/MAPK Signaling by Binding Directly to RAS and Disrupting RAS Clusters”
https://www.sciencedirect.com/science/article/pii/S221112471931527X#sec5

This antibody was used for immunoblotting of RIBA buffer cell lysates obtained from tumor cell lines PFSK-1 and D324.
Dilution: 1:1000
Ref: Spyrou et al., Mol Cancer Ther, 2017 Aug; 16(8): 1705-1716

This antibody is used for evaluating activity of the EGF-receptor pathway after stimulation or inhibition of EGFR in lung cancer cell lines. The antibody is very reliable. We have found that dilution in 5 % BSA is optimal.
Dilution: 1:500

Phospho-Akt (Ser473) rabbit antibody #9271 from Cell Signaling gives one specific band at 60 kDa.

I have used the antibody for western blotting with 20 ug WCE of cultured primary mouse hippocampal neurons and with 20 ug WCE of the human neuroblastoma cell line SH-SY5Y in the dilution 1:1000 and incubation at +4C ON.

The antbody can be reused 2 times.

rabbit Phospho-Akt (cat no 9271) from Cell Signaling works very nicely in WB. Have detected pAKT levels from both cell-lines (MG87 fibroblasts) as well as from embryonic kidney tissue lysates (E14).

Used dilution 1:1000, 5%BSA/TBS/Tween, incubates O/N at +4C