Human S6K1/RPS6KB1 Baculovirus-Insect cells Overexpression Lysate

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Human S6K1/RPS6KB1 Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human S6K1/RPS6KB1 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of S6K1/RPS6KB1 protein (Cat: 10099-H09B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the human RPS6KB1 (P23443-Alpha I) (Met1-Leu525) was fused with the GST tag at the N-terminus.
Molecule Mass
The recombinant human RPS6KB1 consists of 750 amino acids and predicts a molecular mass of 85.4 KDa. The apparent molecular mass of the protein is approximately 96 Kda in SDS-PAGE under reducing conditions due to glycosylation.

Human S6K1/RPS6KB1 Baculovirus-Insect cells Overexpression Lysate: Usage Guide

Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human S6K1/RPS6KB1 Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human p70 S6KA Overexpression Lysate; Human p70(S6K)-alpha Overexpression Lysate; Human p70-alpha Overexpression Lysate; Human p70-S6K Overexpression Lysate; Human PS6K Overexpression Lysate; Human RPS6KB1 Overexpression Lysate; Human S6K Overexpression Lysate; Human S6K-beta-1 Overexpression Lysate; Human S6K1 Overexpression Lysate; Human STK14A Overexpression Lysate

S6K1/RPS6KB1 Background Information

PS6K, also known as RPS6KB1, is a serine/threonine-protein kinase. It belongs to the RSK (ribosomal s6 kinase) family. Members of this family function in signal transduction. PS6K is an isoform of p7 ribosomal S6 kinase (S6K). S6K can be activated by mitogenic stimuli such as growth factors, insulin and cytokines. It phosphorylates the ribosomal protein S6. PS6K also phosphorylates other proteins such as elF4B, eEF2K and SKAR. It is a crucial effector of mTOR(rapamycin) signaling. PS6K is dissociated from the EIF3 complex and activated upon mitogenic stimulation, phosphorylation by the mammalian target of mTOR complex 1 (mTORC1). Its active form then phosphorylates and activates several substrates in the preinitiation complex, including the EIF2B complex and the cap-binding complex component EIF4B. PS6K also functions in cell proliferation, cell growth and cell cycle progression.
Full Name
ribosomal protein S6 kinase, polypeptide 1
References
  • Panasyuk, et al. (2006) Nuclear export of PS6K II is regulated by protein kinase CK2 phosphorylation at Ser-17. J Biol Chem. 281(42):31188-201.
  • Carnevalli L, et al. (2010) PS6K Plays a Critical Role in Early Adipocyte Differentiation. Dev Cell. 18 (5):763-74.
  • Grove JR, et al. (1991) Cloning and expression of two human p70 S6 kinase polypeptides differing only at their amino termini. Mol Cell Biol. 11(11):5541-50.
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