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Pgsk3α β s21 9

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PGSK3α/β S21/9 is a product offered by Cell Signaling Technology. It is a phospho-specific antibody that detects endogenous levels of GSK3α/β when phosphorylated at Ser21/Ser9.

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Lab products found in correlation

Pan akt, by Cell Signaling Technology (2 mentions) Pakt t308, by Cell Signaling Technology (2 mentions) Pakt s473, by Cell Signaling Technology (2 mentions) Recombinant human tgf β1, by Thermo Fisher Scientific (1 mentions) Ps6 s240 244, by Cell Signaling Technology (1 mentions) Recombinant human egf, by Thermo Fisher Scientific (1 mentions) Rad001, by Selleck Chemicals (1 mentions) Antibody against hsc 70, by Santa Cruz Biotechnology (1 mentions) Pakt1 s473, by Cell Signaling Technology (1 mentions) Pakt2 s474, by Cell Signaling Technology (1 mentions) Pher2 y877, by Cell Signaling Technology (1 mentions) Mmp 2, by Cell Signaling Technology (1 mentions) α tubulin, by Merck Group (1 mentions) P jnk t183 y185, by Cell Signaling Technology (1 mentions) P p38 t180 y182, by Cell Signaling Technology (1 mentions) P src y416, by Cell Signaling Technology (1 mentions) P ndrg1 t346, by Cell Signaling Technology (1 mentions) Ps6 s235 236, by Cell Signaling Technology (1 mentions) Alexa fluor 488 donkey anti rabbit, by Thermo Fisher Scientific (1 mentions) Cleaved caspase 3 asp175, by Cell Signaling Technology (1 mentions)

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PGSK3α/β (15 citations)

6 protocols using pgsk3α β s21 9

1

Comprehensive Signaling Pathway Analysis

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Antibodies against AKT1, AKT2, AKT3, panAKT, pAKT S473, pAKT T308, pAKT1 S473, pAKT2 S474, pS6 S240/244, pGSK3α/β S21/9, pHER2 Y877, S100A4, MMP2, RANK, and CTGF, and secondary HRP-linked antibodies against rabbit or mouse IgG were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). Antibody against AKT3 was provided by Millipore (Burlington, MA, USA). Antibody against HSC-70 was purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Antibody against pDDR1/2 Y796/Y740 was provided by R&D systems Inc. (Minneapolis, MN, USA). RAD001 was provided by Selleck Chemicals (Houston, TX, USA) and MK-2206 was obtained from AbMole BioScience Inc. (Houston, TX, USA). Recombinant human TGF-β1 and recombinant human EGF were purchased from PeproTech Inc. (Rocky Hill, NJ, USA).
Hinz N., Baranowsky A., Horn M., Kriegs M., Sibbertsen F., Smit D.J., Clezardin P., Lange T., Schinke T, & Jücker M. (2021). Knockdown of AKT3 Activates HER2 and DDR Kinases in Bone-Seeking Breast Cancer Cells, Promotes Metastasis In Vivo and Attenuates the TGFβ/CTGF Axis. Cells, 10(2), 430.
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2

Comprehensive Antibody Immunoblotting Protocol

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Antibodies against beta-actin (1:5000, catalog no. sc-69879), Pan 14-3-3 (1:10,000, catalog no. sc-1657), and MAP4K5 (KHS, 1:500, catalog no. sc-6429) were purchased from Santa Cruz Biotechnology (Dallas, TX). α-tubulin (1:5000, catalog no. T5168) was purchased from Sigma. SGK1 (1:1000, catalog no. 3272 and 12103), pErk-T202/Y204 (1:2000, catalog no. 4370S), Erk (1:2000, catalog no. 4695S), pJNK-T183/Y185 (1:1000, catalog no. 9252), JNK (1:2000, catalog no. 9251), p-p38-T180/Y182 (1:1000, catalog no. 4511), p38 (1:2000, catalog no. 9212), pYAP-S127 (1:2000, catalog no. 4911), YAP (1:2000, catalog no. 14074), pNDRG1-T346 (1:2000, catalog no. 5482), NDRG1 (1:2000, catalog no. 9485), pS6-S235/236 (1:1000, catalog no. 2211), S6 (1:1000, catalog no. 2217), pGSK3α/β-S21/9 (1:1000, catalog no. 9331), pSrc-Y416 (1:1000, catalog no. 2123), and cleaved Caspase-3 (Asp175) (1:100 for IF, catalog no. 9664) were purchased from Cell Signaling Technology (Danvers, MA). Ki67 (1:200 for IF, catalog no. RM-9106) was purchased from Thermo Fisher. Donkey anti-rabbit Alexa Fluor® 488 (1:500, catalog no. A-21206) was purchased from Life Technologies.
Ma X., Zhang L., Song J., Nguyen E., Lee R.S., Rodgers S.J., Li F., Huang C., Schittenhelm R.B., Chan H., Chheang C., Wu J., Brown K.K., Mitchell C.A., Simpson K.J, & Daly R.J. (2019). Characterization of the Src-regulated kinome identifies SGK1 as a key mediator of Src-induced transformation. Nature Communications, 10, 296.
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3

Immunoblot Analysis of mTOR Pathway in DCs

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Immunoblots were performed as described [27 (link)]. Briefly, DCs were lysed in CelLytic M (Sigma-Aldrich) containing a protease inhibitor cocktail (Mini-Tablets; Roche Diagnostics). Debris was removed by centrifugation and protein separated on a 4–20% SDS-polyacrylamide electrophoresis gel before membrane transfer. Immunoblotting was performed with primary Abs from Cell Signaling Technology, directed against mTOR (#2983), Raptor (#2280), Rictor (#2114), p70S6K (#9202), p-p70S6K (T389) (#9234), Akt (#9272), p-Akt (S473) (#9271), FoxO1 (#2880), p-FoxO1 (S256) (#9461), p-GSK-3α/β (S21/9) (#9327), p-GSK-3β (S9) (#9336), NF-κB p65 (#4764), or p-NF-κB p65 (S563) (#3033). In addition, mAbs to GAPDH or β-actin (Novus Biologicals) were used. After washing, membranes were incubated with horseradish peroxidase-conjugated Abs (Cell Signaling Technology). Band visualization was achieved with SuperSignal West Pico Substrate (Pierce Chemical) and exposure to film. The intensity of individual bands was quantified using ImageJ (NIH) relative to the loading control (β-actin or GAPDH) and represented as relative expression compared to the non-stimulated control band.
Raïch-Regué D., Rosborough B.R., Watson A.R., McGeachy M.J., Turnquist H.R, & Thomson A.W. (2015). mTORC2 deficiency in myeloid DC enhances their allogeneic Th1 and Th17 stimulatory ability after TLR4 ligation in vitro and in vivo. Journal of immunology (Baltimore, Md. : 1950), 194(10), 4767-4776.
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4

mTOR Pathway Protein Modulation

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The following antibodies were purchased from Cell Signaling Technologies: p-4E-BP1 S65 (#9451), p-S6K1 T389 (#9234), S6K1 (#2708), mTOR (#2983), p-TSC2 T1462 (#3611), p-Akt T308 (#13038), p-Akt S473 (#4060), p-ULK S757 (#6888), ULK1 (8054), 4E-BP1 (#9644), ubiquitin (#3933), ATG5 (#12994), Akt1 (#2938), Akt2 (#3063), Akt3 (#14982), pan-Akt (#4691), p-GSK3α/β S21/9 (#9331), GSK3 α/β (5676), p-GRB10 Ser476 (11817), Grb10 (3702), RagA/B (#4357), and p-Tyr (#8954). Antibodies to vinculin (V9264) and GAPDH (G8795) were purchased from Sigma. The antipuromycin antibody clone 12D10 (MABE343) was purchased from EMD-Millipore. Calnexin was purchased from Abcam (ab22595) and LC3B from Novus Biologicals (NB100-2220). Rapamycin (used at a final concentration of 50 nM) and MG132 were purchased from Calbiochem. MK2206 was purchased from Medchem express. GSK690693 and GDC0941 were purchased from Selleck Chemicals. Chloroquine was purchased from Invitrogen; puromycin and cycloheximide were purchased from Sigma–Aldrich.
Buel G.R., Dang H.Q., Asara J.M., Blenis J, & Mutvei A.P. (2022). Prolonged deprivation of arginine or leucine induces PI3K/Akt-dependent reactivation of mTORC1. The Journal of Biological Chemistry, 298(6), 102030.
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5

Signaling Pathways Antibody Detection

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Specific antibodies for pAkt(Thr308)(#2965), pAkt(Ser473)(#4051), Akt(#4691), pGSK3α/β(S21/9)(#9331), GSK3(#5676), p-ERK1/2(T202/Y204)(#4370), ERK1/2(#4695), and PTEN(#9559) were obtained from Cell Signaling Technology (MA). VO-OHpic and Wortmannin were obtained from Sigma Aldrich. Insulin, LatrunculinB, and LY294002 were obtained from Wako Pure Chemical Industries (Japan).
Katsura Y., Kubota H., Kunida K., Kanno A., Kuroda S, & Ozawa T. (2015). An optogenetic system for interrogating the temporal dynamics of Akt. Scientific Reports, 5, 14589.
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6

IHC Assay for pGSK3α/β in NSCLC

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IHC on formalin-fixed and paraffin-embedded human NSCLC tissues collected by the Emory University/Winship Cancer Institute human tissue procurement services core was performed using Cytomation EnVision®+Dual Link System-HPR (DAB+) (Dako North America, Inc., Carpinteria, CA) following the standard manufacturer’s protocol. Tissues were incubated overnight at 4°C with the primary antibody against p-GSK3α/β (S21/9; Cell Signaling Technology, Inc) at 1:75 dilution. p-GSK3α/β staining was scored as negative (< 10% staining) and positive (≥10% staining) staining, respectively.
Koo J., Yue P., Gal A.A., Khuri F.R, & Sun S.Y. (2014). Maintaining glycogen synthase kinase-3 activity is critical for mTOR kinase inhibitors to inhibit cancer cell growth. Cancer research, 74(9), 2555-2568.
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