Pan akt

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Pan-Akt is a laboratory product designed to detect and quantify the total Akt protein levels in various cell and tissue samples. It is a critical tool for researchers studying cell signaling pathways, cell growth, and survival.

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3 protocols using pan akt

Whole Cell Lysate Preparation and Immunoprecipitation

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To prepare whole cell lysates, cells or tissues were lysed in Buffer B as described previously [7 (link)]. For immunoprecipitations, 200 μg whole cell lysate was diluted in IP buffer and immunoprecipitated with 4G10 anti-phosphotyrosine agarose conjugate (05-777, Millipore) as described previously [9 (link)]. Whole cell lysates and immunoprecipitates were processed by WES to separate and visualize cellular proteins according to a standard instrument protocol. Primary antibodies used were: Ron β (#sc-374626, 1:25), GAPDH (#sc-25778, 1:300), PCNA (#sc-56, 1:25) from Santa Cruz Biotechnology; pan AKT (#4691, 1:25), phospho-AKT (#9271, 1:25), S6 (#2217, 1:25), phospho-S6 (#4856, 1:25), phospho-eIF4B (#3591, 1:25), phosphor-AKT1(# 9018, 1:25), phosphor-AKT2 (# 8599, 1:25), AKT1(# 2938, 1:25), AKT2 (# 3063, 1:25), phospho-S6K1 (#9234, 1:25), S6K1 (#2708, 1:25), cleaved PARP (#9541, 1:25), phospho-GSK3α/β (#9331, 1:25), GSK3α/β (#5676, 1:25), Met (#8198, 1:25), Ret (#14556, 1:25) and Axl (#8661, 1:25) from Cell Signaling Technology. Secondary antibodies were included in a Wes Master Kit (PS-MK14, ProteinSimple).

Wang L., Wang L., Cybula M., Drumond-Bock A.L., Moxley K.M, & Bieniasz M. (2020). Multi-kinase targeted therapy as a promising treatment strategy for ovarian tumors expressing sfRon receptor. Genes & Cancer, 11(3-4), 106-121.

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Molecular Signaling in Cisplatin-Induced Injury

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Cisplatin and TGF-β1 (Sigma-Aldrich, St. Louis, MO, USA) were dissolved in PBS and 10 mM Citric Acid (pH 3.0), respectively. Small inhibitors LY294002, SB203580, U0126, SP600125 (SelleckChemicals, Houston, TX, USA) were dissolved in DMSO. Phospho-Akt (Ser473), phospho-Smad3 (Ser423/425), phospho-ERK (Thr202/Tyr204), Erk, phospho-p38 (Thr180/Tyr182), SAPK/JNK, phospho-SAPK/JNK (Thr183/Tyr185), p15^INK4B, p21^Waf1/Cip1, bax, bcl2, and ki67 were purchased from Cell Signaling Technology (Beverly, MA, USA). P38, pan-Akt, and β-Actin were purchased from Santa Cruz Biotechnology Inc (Santa Cruz, CA, USA). Smad3 and c-myc were purchased from Abcam Co Ltd (Cambridge, UK). Horse radish peroxidase (HRP) conjugated secondary anti-rabbit and antimouse IgG antibodies were from Sigma-Aldrich Co., Ltd.

Zhou H.H., Chen L., Liang H.F., Li G.Z., Zhang B.X, & Chen X.P. (2016). Smad3 Sensitizes Hepatocelluar Carcinoma Cells to Cisplatin by Repressing Phosphorylation of AKT. International Journal of Molecular Sciences, 17(4), 610.

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Statins Modulate Insulin Signaling

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Primary adipocytes from the mouse visceral adipose tissue and 3T3-L1, HepG2, and C2C12 cells were pretreated with 10 µM pitavastatin or rosuvastatin for 24 h, serum-starved for 4 h with or without statins, and stimulated with 100 nM insulin for 5 to 15 min. Cell lysates were obtained from each treatment group.
Equal amounts of proteins were electrophoresed on sodium dodecyl sulfate polyacrylamide gels and the separated protein bands were transferred onto polyvinylidene fluoride membranes. After blocking, the membranes were incubated with phospho-Akt serine 473 (#9271, Cell Signaling Technology, Denver, MA, USA), phospho-Akt threonine 308 (#9275), and pan-Akt (#9272) antibodies, followed by treatment with horseradish peroxidase-conjugated anti-rabbit IgG (sc-2030, Santa Cruz Biotechnology, Santa Cruz, CA, USA). The blots were developed using an enhanced chemiluminescence detection kit. Equal loading was verified by re-probing the blot with β-actin (A5441, Sigma, St Louis, MO, USA) antibody.

Cho Y., Lee H., Park H.K., Choe E.Y., Wang H.J., Kim R.H., Kim Y, & Kang E.S. (2020). Differential Diabetogenic Effect of Pitavastatin and Rosuvastatin, in vitro and in vivo. Journal of Atherosclerosis and Thrombosis, 27(5), 429-440.

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