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P ndrg1

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P-NDRG1 is a rabbit monoclonal antibody that detects endogenous levels of NDRG1 protein when phosphorylated at Thr346. NDRG1 is a protein involved in cellular stress response pathways.

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Lab products found in correlation

P akt, by Cell Signaling Technology (3 mentions) P 4ebp1, by Cell Signaling Technology (3 mentions) Ndrg1, by Cell Signaling Technology (2 mentions) P raptor, by Cell Signaling Technology (2 mentions) P ampkα, by Cell Signaling Technology (2 mentions) Anti actin c4, by Abcam (2 mentions) Compound c, by Enzo Life Sciences (2 mentions) Sodium palmitate, by Nu-Chek Prep (2 mentions) P foxo1 3, by Cell Signaling Technology (2 mentions) P acc1 ser79, by Cell Signaling Technology (2 mentions) Ampkα, by Cell Signaling Technology (2 mentions) Anti flag m2, by Merck Group (2 mentions) Acetylated lysine, by Cell Signaling Technology (1 mentions) Synaptophysin, by Agilent Technologies (1 mentions) Trichostatin a tsa, by Merck Group (1 mentions) Gsk650394, by Bio-Techne (1 mentions) P pkcα, by Santa Cruz Biotechnology (1 mentions) 5 methylcytosine 5mc, by Cell Signaling Technology (1 mentions) Nestin, by Merck Group (1 mentions) Gsk126, by MedChemExpress (1 mentions)

6 protocols using p ndrg1

1

Compound C and Sodium Palmitate Signaling Pathway Analysis

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Compound C was obtained from Enzo Life Sciences. Sodium Palmitate is from NU-CHEK Prep Inc. For western blotting, anti-Flag M2 (RRID:AB_259529) antibody was purchased from Sigma, antibodies against SGK1 (RRID:AB_2687476), SGK2 (RRID:AB_10828732), SGK3 (RRID:AB_10949507), HSP90 (RRID:AB_2233307), Akt (RRID:AB_915783), p-Akt (Thr308) (RRID:AB_2255933), p-Akt (Ser473) (RRID:AB_2315049), NDRG1 (RRID: AB_11140640), p-NDRG1(Thr346) (RRID:AB_10693451), FoxO1 (RRID:AB_2106495), p-FoxO1/3 (Thr24/32) (RRID:AB_2106814), S6K (RRID:AB_390722), p-S6K (Thr389) (RRID:AB_2269803), S6 (RRID:AB_331355), p-S6 (S240/244) (RRID:AB_10694233), 4EBP1 (RRID:AB_2097841), p-4EBP1 (Thr37/46) (RRID:AB_560835), p-4EBP1 (Ser65) (RRID:AB_330947), AMPKα (RRID:AB_10624867), p-AMPKα (Thr172) (RRID:AB_331250), p-AMPKα(Ser485/491) (RRID:AB_331250), ACC1 (RRID:AB_2219397), p-ACC1 (Ser79) (RRID:AB_330337), Raptor (RRID:AB_561245)and p-Raptor (Ser792) (RRID:AB_2249475) were obtained from Cell Signaling Technology. Anti-actin (C4) was obtained from Abcam.
Zhou B., Zhang Y., Li S., Wu L., Fejes-Toth G., Naray-Fejes-Toth A, & Soukas A.A. (2021). Serum- and glucocorticoid-induced kinase drives hepatic insulin resistance by directly inhibiting AMP-activated protein kinase. Cell reports, 37(1), 109785.
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2

Protein Expression Profiling in Cells

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Cell Signaling (Beverly, MA) antibodies: EGF receptor variant III (EGFRvIII) (Cat# 64952), p-Akt (S473; Cat# 4060), p-NDRG1 (T346; Cat# 5482), Rictor (Cat# 2114), 5-methylcytosine (5-mC) (Cat# 28692), DNMT3A (Cat #3598), acetylated-lysine (Cat# 9441), H3 p.K27me2 (Cat# 9755), H3 p.K27me3 (Cat# 9733), Histone H3 (Cat# 4499), EZH2 (Cat# 5246), FAK (Cat# 13009), p-FAK (Y397; Cat# 8556), β-actin (Cat# 3700), GAPDH (Cat# 5174), HRP-linked anti-rabbit IgG (Cat# 7074) and HRP-linked anti-mouse IgG (Cat# 7076). Santa Cruz (Dallas, TX) antibodies: p-PKC α (S657; Cat# sc-377565). GeneTex (Irvine, CA) antibodies: 5-mC (Cat# GT4111). Thermo Fisher antibodies: GRIA1 (Cat# PA5-95207). DAKO (Glostrup, Denmark) antibodies: Synaptophysin (Cat# M731529). Millipore (Burlington, MA) antibodies: Nestin (Cat# MAB5326).
Reagents used are sodium acetate (Sigma; Cat # S5636), Trichostatin A (TSA) (Sigma; Cat# T1952), PP242 (Cayman Chemical, Ann Arbor, MI; Cat# 13643), Akti-1/2 (Calbiochem, La Jolla, CA; Cat# 124018), Bisindolylmaleimide I (Bis-I) (Santa Cruz; Cat# sc-24003), GSK 650394 (Tocris Bioscience, Bristol, UK; Cat# 3572/10), GSKJ4 (Sigma; Cat# T1952), GSK126 (MedChem Express, Monmouth Junction, NJ; Cat# HY-13470), GSK2256098 (Selleck Biotech, Kanagawa, Japan; Cat# S8523) and Philanthotoxin-7,4 (PhTx-74) (Abcam, Cambridge, UK; Cat# ab120257).
Harachi M., Masui K., Shimizu E., Murakami K., Onizuka H., Muragaki Y., Kawamata T., Nakayama H., Miyata M., Komori T., Cavenee W.K., Mischel P.S., Kurata A, & Shibata N. (2024). DNA hypomethylator phenotype reprograms glutamatergic network in receptor tyrosine kinase gene-mutated glioblastoma. Acta Neuropathologica Communications, 12, 40.
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3

Immunohistochemical Profiling of Cell Signaling

Cited 1 time
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Immunohistochemistry (IHC) for all targets was performed using standard protocols on the automated immunohistochemistry staining system Discovery XT (Roche/Ventana, Tucson, Arizona, USA) and the LEICA BOND-III automated stainer (Leica, Wetzlar, Germany) respectively. The following antibodies were used: PD-L1 (Cell Signaling, Boston, U.S.A.), p-S6K1 (Cell Signaling, Boston, U.S.A.), p-4EBP1 (Cell Signaling, Boston, U.S.A.), p-RPS6 (Ser 235/236 and Ser240/244, Cell Signaling, Boston, U.S.A.), p-PRAS40 (Cell Signaling, Boston, U.S.A.), p-NDRG1 (Cell Signaling, Boston, U.S.A.), p-mTOR (S2448, Cell Signaling, Boston, U.S.A.), IDH1_R132H (Dianova, Eching, Germany) and BRAF V600E (DCS, Hamburg, Germany) [27 (link)].
Luger A.L., König S., Samp P.F., Urban H., Divé I., Burger M.C., Voss M., Franz K., Fokas E., Filipski K., Demes M.C., Stenzinger A., Sahm F., Reuss D.E., Harter P.N., Wagner S., Hattingen E., Wichert J., Lapa C., Fröhling S., Steinbach J.P, & Ronellenfitsch M.W. (2022). Molecular matched targeted therapies for primary brain tumors—a single center retrospective analysis. Journal of Neuro-Oncology, 159(2), 243-259.
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4

Compound C and Sodium Palmitate Signaling Pathway Analysis

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Compound C was obtained from Enzo Life Sciences. Sodium Palmitate is from NU-CHEK Prep Inc. For western blotting, anti-Flag M2 (RRID:AB_259529) antibody was purchased from Sigma, antibodies against SGK1 (RRID:AB_2687476), SGK2 (RRID:AB_10828732), SGK3 (RRID:AB_10949507), HSP90 (RRID:AB_2233307), Akt (RRID:AB_915783), p-Akt (Thr308) (RRID:AB_2255933), p-Akt (Ser473) (RRID:AB_2315049), NDRG1 (RRID: AB_11140640), p-NDRG1(Thr346) (RRID:AB_10693451), FoxO1 (RRID:AB_2106495), p-FoxO1/3 (Thr24/32) (RRID:AB_2106814), S6K (RRID:AB_390722), p-S6K (Thr389) (RRID:AB_2269803), S6 (RRID:AB_331355), p-S6 (S240/244) (RRID:AB_10694233), 4EBP1 (RRID:AB_2097841), p-4EBP1 (Thr37/46) (RRID:AB_560835), p-4EBP1 (Ser65) (RRID:AB_330947), AMPKα (RRID:AB_10624867), p-AMPKα (Thr172) (RRID:AB_331250), p-AMPKα(Ser485/491) (RRID:AB_331250), ACC1 (RRID:AB_2219397), p-ACC1 (Ser79) (RRID:AB_330337), Raptor (RRID:AB_561245)and p-Raptor (Ser792) (RRID:AB_2249475) were obtained from Cell Signaling Technology. Anti-actin (C4) was obtained from Abcam.
Zhou B., Zhang Y., Li S., Wu L., Fejes-Toth G., Naray-Fejes-Toth A, & Soukas A.A. (2021). Serum- and glucocorticoid-induced kinase drives hepatic insulin resistance by directly inhibiting AMP-activated protein kinase. Cell reports, 37(1), 109785.
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5

Examining Phosphorylation Signaling Pathways

Cited 1 time
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The following antibodies were purchased from Cell Signaling Technologies: pNDRG1, pPDK1, pAKT308, pAKT473, pGSK3β (Ser 9), pPRAS40 (Thr 246), pFOXO3a (Thr 32), pERK1/2, AKT, FOXO3a, pP70S6K (Thr 389), pS6K (Ser 235/236), PRAS40, GSK3b, ERK1/2, SGK3, cyclin D1, PTEN, P70S6K, and S6. Antibodies against β-actin, SGK1, PKC, and tubulin were purchased from Santa Cruz Biotechnology. 4-Hydroxytamoxifen and antibodies against S100 were purchased from Sigma. Synthesis of the PDK1 inhibitor GSK2334470 was performed as previously described (5 (link)).
Scortegagna M., Lau E., Zhang T., Feng Y., Sereduk C., Yin H., De S.K., Meeth K., Platt J.T., Langdon C.G., Halaban R., Pellecchia M., Davies M.A., Brown K., Stern D.F., Bosenberg M, & Ronai Z.A. (2015). PDK1 and SGK3 contribute to the growth of BRAF mutant melanomas and are potential therapeutic targets. Cancer research, 75(7), 1399-1412.
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6

Prostate Cancer Cell Proliferation Assay

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For Western Blotting, 2 million LNCaP (ATTC) cells were plated on 60 mm dishes in RPMI-1640 containing 10% FBS and 1% pen-strep antibiotic. After 24 h plating cell media was replaced with RPMI-1640 containing 10% charcoal stripped fetal bovine serum to decrease androgen interference from the media. Cells were either transfected with SGK-DN or treated with drug (500 nM–100 μM and treated with the synthetic androgen analog R1881 (1 nM). Cells were treated at 0, 12 and 24 h and harvested 32 h after first treatment. Proteins were separated by gel electrophoresis using 50 μg of protein per well. P-NDRG1 (Cell Signaling) was normalized to GAPDH and data presented as fold change from control. For CyQUANT cell proliferation assay (Life Technologies), LNCaP cells were plated at 25,000 cells/well in a 96 well plate in plating media. 24 h after plating, media was changed to RPMI-1640 containing 10% charcoal stripped fetal bovine serum. Cells were treated with either 5377051, R1881 (1 nM) or both every 24 h for 72 h. Cell proliferation was measured using the manufacturer’s protocol.
Bezzerides V.J., Zhang A., Xiao L., Simonson B., Khedkar S.A., Baba S., Ottaviano F., Lynch S., Hessler K., Rigby A.C., Milan D., Das S, & Rosenzweig A. (2017). Inhibition of serum and glucocorticoid regulated kinase-1 as novel therapy for cardiac arrhythmia disorders. Scientific Reports, 7, 346.
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