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Sphingosine Kinase 1 Cell Lysate | SPHK1 cell lysate

Human Sphingosine Kinase 1 Baculovirus-Insect cells Overexpression Lysate

Gene Names
SPHK1; SPHK
Applications
Western Blot
Synonyms
Sphingosine Kinase 1; Human Sphingosine Kinase 1 Baculovirus-Insect cells Overexpression Lysate; Human SPHK1/Sphingosine Kinase 1 Insect Cell Lysate (WB positive control); sphingosine kinase 1; Human SPHK Overexpression Lysate; Human SPHK1 Overexpression Lysate; Human SPK Overexpression Lysate; SPHK1 cell lysate
Ordering
For Research Use Only!
Host
Baculovirus-Insect cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading 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.
Sequence Positions
621aa
Sequence
Met1-Leu384
Applicable Applications for SPHK1 cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Human
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 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the human SPHK1 (Q9NYA1-1) (Met1-Leu384) was expressed the N-terminal polyhistidine-tagged GST tag at the N-terminus.
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. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for SPHK1 cell lysate
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.

This Human Sphingosine Kinase 1 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Sphingosine Kinase 1 protein from the overexpression lysate was verified.
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.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
UniProt Accession #
Molecular Weight
384
NCBI Official Full Name
Sphingosine kinase 1
NCBI Official Synonym Full Names
sphingosine kinase 1
NCBI Official Symbol
SPHK1
NCBI Official Synonym Symbols
SPHK
NCBI Protein Information
sphingosine kinase 1; SK 1; SPK 1
UniProt Protein Name
Sphingosine kinase 1
Protein Family
UniProt Gene Name
SPHK1
UniProt Synonym Gene Names
SPHK; SPK; SK 1; SPK 1
UniProt Entry Name
SPHK1_HUMAN

NCBI Description

The protein encoded by this gene catalyzes the phosphorylation of sphingosine to form sphingosine-1-phosphate (S1P), a lipid mediator with both intra- and extracellular functions. Intracellularly, S1P regulates proliferation and survival, and extracellularly, it is a ligand for cell surface G protein-coupled receptors. This protein, and its product S1P, play a key role in TNF-alpha signaling and the NF-kappa-B activation pathway important in inflammatory, antiapoptotic, and immune processes. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Sep 2011]

Uniprot Description

SPHK1: Catalyzes the phosphorylation of sphingosine to form sphingosine 1-phosphate (SPP), a lipid mediator with both intra- and extracellular functions. Also acts on D-erythro-sphingosine and to a lesser extent sphinganine, but not other lipids, such as D,L-threo-dihydrosphingosine, N,N-dimethylsphingosine, diacylglycerol, ceramide, or phosphatidylinositol. Interacts with ACY1. Binds to calmodulin. Interacts with SPHKAP. Widely expressed with highest levels in adult liver, kidney, heart and skeletal muscle. 2 isoforms of the human protein are produced by alternative splicing.

Protein type: Lipid Metabolism - sphingolipid; EC 2.7.1.91; Kinase, lipid

Chromosomal Location of Human Ortholog: 17q25.2

Cellular Component: synaptic vesicle; axon; cytoplasm; plasma membrane; nucleus; cytosol

Molecular Function: D-erythro-sphingosine kinase activity; calmodulin binding; protein binding; DNA binding; protein phosphatase 2A binding; sphinganine kinase activity; magnesium ion binding; diacylglycerol kinase activity; ATP binding; NAD+ kinase activity

Biological Process: protein folding; positive regulation of NF-kappaB import into nucleus; female pregnancy; positive regulation of mitotic cell cycle; signal transduction; positive regulation of neurotransmitter secretion; activation of NF-kappaB transcription factor; 'de novo' posttranslational protein folding; response to magnesium ion; positive regulation of fibroblast proliferation; regulation of interleukin-1 beta production; protein kinase C activation; positive regulation of smooth muscle contraction; inflammatory response; blood vessel development; sphingolipid metabolic process; sphingolipid biosynthetic process; cyclooxygenase pathway; calcium-mediated signaling; sphingosine metabolic process; cellular response to starvation; lipid phosphorylation; positive regulation of cell growth; response to ATP; positive regulation of angiogenesis; cellular protein metabolic process; positive regulation of protein ubiquitination; brain development; positive regulation of protein amino acid phosphorylation; response to progesterone stimulus; vascular endothelial growth factor receptor signaling pathway; sphingoid catabolic process; negative regulation of apoptosis; positive regulation of cell migration; response to amine stimulus

Research Articles on SPHK1

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Product Notes

The SPHK1 sphk1 (Catalog #AAA8115543) is a Cell Lysate produced from Baculovirus-Insect cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 621aa. AAA Biotech's Sphingosine Kinase 1 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the SPHK1 sphk1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Met1-Leu38 4. It is sometimes possible for the material contained within the vial of "Sphingosine Kinase 1, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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