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Human SPHK1 / Sphingosine Kinase 1 Insect Cell Lysate (WB positive control)

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Human SPHK1 Transfected / Overexpression Cell Lysate Product Information
Expressed Host:Baculovirus-Insect cells
Product Description:Baculovirus-Insect Cell lysate that Human SPHK1 / Sphingosine Kinase 1 transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
Sequence information:A DNA sequence encoding the human SPHK1 (Q9NYA1-1)(Met1-Leu384) was fused with two additional amino acids (Gly & Pro) at the N-terminus.
Predicted N Terminal:Gly
Molecule Mass:The recombinant human SPHK1 consists of 386 amino acids and has a calculated molecular mass of 42.7 kDa. The recombinant protein migrates as an approximately 46 kDa band in SDS-PAGE under reducing conditions.
Human SPHK1 Transfected / Overexpression Cell Lysate Usage Guide
Preparation Method:Cell lysate was prepared by homogenization 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.
Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.
Stability:Samples are stable for up to twelve months from date of receipt.
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. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
Storage Buffer:1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Storage Instruction:Store at 4℃. After re-dissolution, aliquot and store at -80℃.
Application notes:Western blot (WB): Use at an assay dependent dilution.
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
SPHK1 / Sphingosine Kinase 1 Background

SPHK1, also known as sphingosine Kinase 1, catalyzes the phosphorylation of sphingosine to form sphingosine-1-phosphate (S1P). S1P exhibits a broad spectrum of biological activities including cell proliferation, survival, migration, cytoskeletal organization, and morphogenesis. It is a ligand for cell surface G protein-coupled receptors. SPHK 1 is a potential therapeutic target for the control of cancer and inflammation. SPHK1 plays a key role in TNF-alpha signaling and the NF-kappa-B activation pathway important in inflammatory, antiapoptotic, and immune processes.

Human SPHK1 / Sphingosine Kinase 1 References
  • Kohama T. et al., 1998, J Biol Chem. 273 (37): 23722-8.
  • Xia P. et al., 2002, J Biol Chem. 277 (10): 7996-8003.
  • Tsukahara T. et al., 2002, Tanpakushitsu Kakusan Koso. 47 (4): 509-13.
  • Size / Price
    Catalog: 15679-HNCBL-300
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