Phospho pras40 t246

Manufactured by Cell Signaling Technology

Sourced in United States, Germany

Phospho-PRAS40 (T246) is a laboratory reagent used to detect the phosphorylation of PRAS40 (proline-rich Akt substrate of 40 kDa) at threonine residue 246. It is typically used in Western blotting and other protein analysis techniques to study the activation of the PI3K/Akt signaling pathway.

Automatically generated - may contain errors

Lab products found in correlation

3 protocols using phospho pras40 t246

Reagents for P2X7 Receptor Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Carbenoxolone (CBX), BAPTA-AM and thapsigargin (TG) were purchased from Tocris Bioscience (Ellisville, MO). APT102, the recombinant form of soluble CD39, was kindly provided by Dr. Ridong Chen at APT Therapeutics Inc. (St. Louis, MO). All other chemicals and cell culture media were from Sigma-Aldrich (St. Louis, MO) and other culture reagents from Life Technologies (Carlsbad, CA), unless otherwise stated.
Rabbit anti-mouse P2X7 antibody (#APR-004) was obtained from Alomone labs (Jerusalem, Israel); beta-actin (AC-15, #ab6276) from Abcam (Cambridge, MA); Flotillin-2 (B6, #sc-28320) from Santa Cruz Biotechnology, Inc. (Dallas, TX); Caveolin-1 (7C8, #NB100-615) from Novus Biologicals (Littleton, CO); phospho-AKT (S473) (#9271), phospho-PRAS40 (T246) (#2997), phospho-S6K (T389) (#9205), phospho-S6 (S235/236) (#2211) from Cell Signaling Technology (Danvers, MA); HRP-conjugated goat anti-mouse (#31438), donkey anti-rabbit (#31458) and mouse anti-goat IgG (#31400) and the SuperSignal West Femto Maximum Sensitivity Substrate reagents (#PI-34096) were from Thermo Scientific (Rockford, IL).

de Andrade Mello P., Bian S., Savio L.E., Zhang H., Zhang J., Junger W., Wink M.R., Lenz G., Buffon A., Wu Y, & Robson S.C. (2017). Hyperthermia and associated changes in membrane fluidity potentiate P2X7 activation to promote tumor cell death. Oncotarget, 8(40), 67254-67268.

+ Open protocol

+ Expand

Western Blot Analysis of Cellular Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed in cell lysis buffer (20 mM Tris, pH 7.5, 135 mM NaCl, 5% [v/v] glycerol, 50 mM NaF, 0.1% [v/v] Triton X-100, plus protease inhibitors), centrifuged and protein quantified using Bradford reagent (Sigma-Aldrich). Samples were made up in NuPAGE LDS sample buffer (Life Technologies). Western blotting was performed as previously described [28 (link)]. Blots shown are representative of 3 independent experiments. Anti-β-actin (#4967), phospho-TSC2 S1387 (#5584), total TSC2 (#3990), IRE1α (3294S), phospho-S6K1 T389 (#9205), total S6K1 (#9202), phospho-rpS6 S235/236 (#2211), total rpS6 (#2217), phospho-4E-BP1 S65 (#9451), total 4E-BP1 (#9644), phospho-ACC S79 (#3661), total ACC (#3676), PARP (#9542), caspase 3 (#9662), ATF4 (#11815), phospho-FOXO3a (#9466S), total FOXO3a (#9467), phospho-PRAS40 T246 (#2997), total PRAS40 (#2691) and phospho-Bad S136 (#4366) antibodies were from Cell Signaling Technology (Danvers, MA, USA). GADD34 antibody (10449-1-AP) was from Proteintech (Manchester, UK).

Dunlop E.A., Johnson C.E., Wiltshire M., Errington R.J, & Tee A.R. (2017). Targeting protein homeostasis with nelfinavir/salinomycin dual therapy effectively induces death of mTORC1 hyperactive cells. Oncotarget, 8(30), 48711-48724.

+ Open protocol

+ Expand

Evaluation of mTOR and PI3K/Akt Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies against phospho-mTOR (S2448, Cat# 2971), mTOR, phospho-S6 (S235/236, Cat# 4856), S6, phospho-Akt1 (S473, Cat# 9271), phospho-Akt1 (T308, Cat# 9275), phospho-GSK-3α/β (S21/9, Cat# 8566), GSK-3α/β (Cat# 5676), phospho-PRAS40 (T246, Cat# 2997), PRAS40 (Cat# 2691), P-Akt2 (S474, Cat# 8599), Akt2 (Cat# 5239) and GAPDH (Cat# 2118) were purchased from Cell Signaling Technology (Frankfurt, Germany). Rapamycin was obtained from Enzo Life Science (Loerrach, Germany). The PI3K inhibitor LY294002 was purchased from Calbiochem (Schwalbach, Germany). The Akt inhibitor MK2206 was purchased from Selleck Chemicals (Munich, Germany). SMART pool siRNA against AKT1 (Cat. No. M-0030000-03) and AKT2 (Cat. No. M-003001-02), as well as a non-targeting siRNA (Cat. No. D-001810-10) were purchased from Thermo Scientific Dharmacon (Bonn, Germany). Anti-Akt1 antibody, anti-phospho-histone H2AX (S139) antibody and the Alexa Fluor 488 conjugated secondary antibody have been previously described [16 (link), 23 (link)].

Holler M., Grottke A., Mueck K., Manes J., Jücker M., Rodemann H.P, & Toulany M. (2016). Dual Targeting of Akt and mTORC1 Impairs Repair of DNA Double-Strand Breaks and Increases Radiation Sensitivity of Human Tumor Cells. PLoS ONE, 11(5), e0154745.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!