General description
Sphingosine kinase 1 (UniProt: Q9NYA1; also known as SK 1, SPK 1, EC:2.7.1.91, Acetyltransferase SPHK1, EC:2.3.1.-) is encoded by the SPHK1 (also known as SK1, SPHK, SPK) gene (Gene ID: 8877) in human. SPHK1 is a crucial enzyme that catalyzes the phosphorylation of sphingosine to produce sphingosine-1-phosphate (S1P), a bioactive lipid involved in various cellular processes, including cell proliferation, survival, and migration. SPHK1 is predominantly expressed in the cytoplasm, with functional nuclear export signals facilitating its translocation to the plasma membrane, particularly in response to various stimuli such as growth factors. Its expression profile reveals that its widely expressed with highest levels in adult liver, kidney, heart and skeletal muscle. Studies revealed that SphK1a was ubiquitously expressed in all cancer cells and tissues tested, and in contrast, SphK1b was only expressed in selective cell types in breast, prostate, and lung cancer. SPHK1 is characterized by a two-domain architecture, which houses a catalytic site located between the domains and a hydrophobic lipid-binding pocket within the C-terminal domain. The N-terminal has DAG-kinase catalytic domain (aa 12-159) and 2 nuclear export signal 1 & 2 motif′s. SPHK1 shares two large conserved regions with SPHK2. Three Isoforms of SPHK1 have been described that are produced by alternative splicing. The aberrant localization and overexpression of SPHK1 have been associated with several diseases, including cancer. SPHK1′s role in astrocytomas has been highlighted, with studies indicating that its upregulation correlates with disease progression and poor patient prognosis. This ZooMAb® recombinant monoclonal antibody, generated by our proprietary technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Chen, H., et al. (2022). J. Oncol. 10(1155); 250407; Wang, Z., et al. (2013). Structure. 21(5):798-809).
ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them.
Immunogen
KLH-conjugated linear peptide corresponding to 20 amino acids from the C-terminal region of human SPHK1.
Application
Quality Control Testing
Evaluated by Western Blotting in HepG2 cell lysate.
Western Blotting Analysis: A 1:1,000 dilution of this antibody detected SPHK1 in HepG2 cell lysate.
Tested Applications
Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected SPHK1 in HT-1080 cell lysate.
Affinity Binding Assay: A representative lot of this antibody bound SPHK1 peptide with at least one thousand-fold (1,000X) higher affinity than with non-specific control peptide.
Immunoprecipitation Analysis: 2 µg from a representative lot immunoprecipitated SPHK1 in HepG2 cell lysate.
Note: Actual optimal working dilutions must be determined by end user as specimens and experimental conditions may vary with the end user.
Biochem/physiol Actions
Clone 2H5 is a ZooMAb® recombinant rabbit monoclonal antibody that specifically detects SPHK1. It targets an epitope within 20 amino acids from the C-terminal region.
Physical form
Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.
Preparation Note
300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.
Legal Information
ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.