Akt 40d4

Manufactured by Cell Signaling Technology

AKT (40D4) is a mouse monoclonal antibody that specifically recognizes the AKT protein. AKT is a serine/threonine protein kinase that plays a crucial role in the regulation of various cellular processes, including cell proliferation, survival, and metabolism.

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Lab products found in correlation

Erk1 2, by Cell Signaling Technology (1 mentions) Phospho erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions) Hsp90 f 8, by Santa Cruz Biotechnology (1 mentions) P erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions) Immobilon western hrp substrate, by Merck Group (1 mentions) Protein lysis buffer, by Merck Group (1 mentions) Pakt ser473 d9e, by Cell Signaling Technology (1 mentions) Protease and phosphatase inhibitor cocktail, by Merck Group (1 mentions) Bis tris gel, by Thermo Fisher Scientific (1 mentions) Ps6 ser235 236, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Akt-pan (40D4) (7 citations) Anti-Akt (40D4; (3 citations) AKT (pan; 40D4) mouse mAb (3 citations) Total Akt (40D4, (2 citations)

2 protocols using akt 40d4

Activation of T cell signaling

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Feeder-expanded T cells were starved for 4 h in PBS/0.5% human serum. Samples were placed on ice and PHA stimulation mix was added still on ice. Samples were placed on a 37 °C thermoshaker and at the indicated time points, ice-cold PBS was added and respective samples were placed on ice immediately. Cells were then lysed in RIPA buffer as above, and resolved by SDS-PAGE Western blot. The following primary antibodies were used: HSP90 (F-8, Santa Cruz), AKT (40D4, Cell Signaling), phospho-AKT (Ser473, D94, Cell Signaling), ERK1/2 (137F5, Cell Signaling), phospho-ERK1/2 (Thr202/Tyr204, Cell Signaling).

Serwas N.K., Hoeger B., Ardy R.C., Stulz S.V., Sui Z., Memaran N., Meeths M., Krolo A., Yüce Petronczki Ö., Pfajfer L., Hou T.Z., Halliday N., Santos-Valente E., Kalinichenko A., Kennedy A., Mace E.M., Mukherjee M., Tesi B., Schrempf A., Pickl W.F., Loizou J.I., Kain R., Bidmon-Fliegenschnee B., Schickel J.N., Glauzy S., Huemer J., Garncarz W., Salzer E., Pierides I., Bilic I., Thiel J., Priftakis P., Banerjee P.P., Förster-Waldl E., Medgyesi D., Huber W.D., Orange J.S., Meffre E., Sansom D.M., Bryceson Y.T., Altman A, & Boztug K. (2019). Human DEF6 deficiency underlies an immunodeficiency syndrome with systemic autoimmunity and aberrant CTLA-4 homeostasis. Nature Communications, 10, 3106.

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Western Blot Analysis of Signaling Proteins

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Cells were lysed using protein lysis buffer (Sigma) with protease and phosphatase inhibitor cocktails (Sigma). Protein concentration was determined using a BCA protein assay kit (Pierce). 15-20μg of protein was separated on a 4-12% Bis-Tris gel (Life Technologies) and transferred to PVDF membranes. Protein expression was detected by Western blotting using the following primary antibodies against: S6 (54D2, Cell Signalling), p-S6 (Ser235/236) (2211, Cell Signalling), Erk1/2 (3A7, Cell Signalling), p-Erk1/2 (Thr202/Tyr204) (9101, Cell Signalling), Akt (40D4, Cell Signalling), p-Akt (Ser473) (D9E, Cell Signalling), and Vinculin (VIN-11-5, Sigma). Primary antibodies were detected with HRP-conjugated anti-rabbit or anti-mouse IgG and visualised with Immobilon Western HRP substrate (Merck).

Boumelha J., de Carné Trécesson S., Law E.K., Romero-Clavijo P., Coelho M.A., Ng K., Mugarza E., Moore C., Rana S., Caswell D.R., Murillo M., Hancock D.C., Argyris P.P., Brown W.L., Durfee C., Larson L.K., Vogel R.I., Suárez-Bonnet A., Priestnall S.L., East P., Ross S.J., Kassiotis G., Molina-Arcas M., Swanton C., Harris R, & Downward J. (2022). An immunogenic model of KRAS-mutant lung cancer enables evaluation of targeted therapy and immunotherapy combinations. Cancer research, 82(19), 3435-3448.

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