Anti ppdk1 ser241

Manufactured by Cell Signaling Technology

Sourced in United States

The Anti-pPDK1 Ser241 is a lab equipment product from Cell Signaling Technology that detects the phosphorylation of PDK1 at serine 241.

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3 protocols using anti ppdk1 ser241

Western Blot Analysis of Phosphorylation

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Fresh frozen tissues were lysed with RIPA buffer (50 mM NaCl, 1.0% IGEPAL® CA-630, 0.5% sodium deoxycholate, 0.1% SDS, and 50 mM Tris, pH 8.0, 10 μg/mL aprotinin, 1 mM phenylmethanesulfonyl fluoride (PMSF), 1 mM Na₂VO₄, 10 μg/mL pepstatin, 10 μg/mL leupeptin). Tissue lysates were separated by electrophoresis in SDS-polyacrylamide gels and transferred to nitrocellulose membranes. After blocking, membranes were hybridized with primary antibodies. Subsequently, membranes were incubated with horseradish peroxidase conjugated secondary antibodies and signals were detected with enhanced chemiluminescence. The following antibodies were used : anti-pPDK1 Ser241 (Cell Signaling, #3061), anti-PDK1 (Cell Signaling, #3062), anti-pS6 Ser235/236 (Cell Signaling, #4856), anti-S6 (Cell Signaling, #2217), beta actin-HRP (Sigma, #A3854).

Cho H., Herzka T., Zheng W., Qi J., Wilkinson J.E., Bradner J.E., Robinson B.D., Castillo-Martin M., Cordon-Cardo C, & Trotman L.C. (2014). RapidCaP, a novel GEM model for analysis and therapy of metastatic prostate cancer reveals Myc as a driver of Pten-mutant metastasis. Cancer discovery, 4(3), 318-333.

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Signaling Pathway Protein Analysis

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The following reagents and materials were used: anti-Bcl-xL (ref. 2764), anti-pAkt (Ser473) (ref. 4060), anti-Akt (ref. 4685), anti-pMEK1/2 (Ser217/221) (ref. 9154), anti-MEK1/2 (ref. 9126), anti-pERK1/2 (Thr202/Tyr204) (ref. 4370), anti-ERK1/2 (ref. 9102), anti-pPKA (Thr197) (ref. 5661), anti-PKA (ref. 4782), anti-pSEK1/MKK4 (Ser257/Thr261) (ref. 9156), anti-SEK1/MKK4 (ref. 9152), anti pSAPK/JNK (Thr183/Tyr185) (ref. 9255), anti pSAPK/JNK (ref. 9252S), anti p-p38 MAPK (Thr180/Tyr182) (ref. 4511), anti p38 MAPK (ref. 9212), anti-pCAMKII (Thr286) (ref. 12,716), anti-CAMKII (ref.11945), anti-pPDK1 (ser241) (ref. 3061), anti-PDK1 (ref. 3062) and anti-pPKC-pan (ref. 9379) were purchased from Cell signaling. Anti-GFAP (ref. ab7260) was purchased from Abcam. Anti PKC-pan (ref. SAB4502356) was from Sigma Aldrich. Electrophoresis reagents were purchased from Biorad and trypsin from Promega.

Lachén-Montes M., Mendizuri N., Ausin K., Andrés-Benito P., Ferrer I., Fernández-Irigoyen J, & Santamaría E. (2020). Amyotrophic Lateral Sclerosis Is Accompanied by Protein Derangements in the Olfactory Bulb-Tract Axis. International Journal of Molecular Sciences, 21(21), 8311.

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Tanshinone Bioactivity Evaluation Protocol

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Bovine serum albumin, sulforhodamine B (SRB), propidium iodide (PI), anti-β-actin antibody, dimethyl sulfoxide, and ribonuclease A (RNase A) were purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA). Dulbecco’s Modified Eagle Medium (DMEM), FBS, and antibiotic-antimycotic solution were purchased from HyClone Laboratories, Inc. (South Logan, UT, USA). Anti-cyclin A, anti-CDK4, anti-CDK2, anti-cMyc, and anti-p21 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-p-Rb (S807/811), anti-Rb, anti-p-Akt (Ser473), anti-Akt, anti-p-mTOR (Ser2448), anti-mTOR, anti-p-p70S6K1 (Thr389), anti-p70S6K1, anti-p-4EBP1 (Thr37/46), anti-4EBP1, anti-p-PDK1 (Ser241), anti-p-AMPK (Thr172), and anti-AMPK were obtained from Cell Signaling (Danvers, MA, USA). Anti-cyclin D1 and anti-Cyclin E were purchased from BD Biosciences (San Jose, CA, USA). Four tanshinones (tanshinone I, tanshinone IIA, cryptotanshinone, and DHTS; Fig. 1) were isolated by Dr. Seung Ho Lee (Yeungnam University, Korea) from the roots of S. miltiorrhiza Bunge (Labiatae) and were provided through the Research Center for Standardization of Herbal Medicines in Korea.

Hong J.Y., Park S.H., Park H.J, & Lee S.K. (2018). Anti-proliferative Effect of 15,16-Dihydrotanshinone I Through Cell Cycle Arrest and the Regulation of AMP-activated Protein Kinase/Akt/mTOR and Mitogen-activated Protein Kinase Signaling Pathway in Human Hepatocellular Carcinoma Cells. Journal of Cancer Prevention, 23(2), 63-69.

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