Human Sphingosine Kinase 1/SPHK1 Antibody Summary
Accession # NP_068807
This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human Sphingosine Kinase 1/SPHK1 Monoclonal Antibody (Catalog # MAB5536).
This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human Sphingosine Kinase 1/SPHK1 DuoSet ELISA Kit (Catalog # DY5536) for convenient development of a sandwich ELISA.
5536-SK), see our available Western blot detection antibodies
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human Sphingosine Kinase 1/SPHK1 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line. PVDF Membrane was probed with 0.5 µg/mL of Sheep Anti-Human Sphingosine Kinase 1/SPHK1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5536) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Sphingosine Kinase 1/SPHK1 at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Sphingosine Kinase 1/SPHK1 in HepG2 Human Cell Line. Sphingosine Kinase 1/SPHK1 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Sheep Anti-Human Sphingosine Kinase 1/SPHK1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5536) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Sphingosine Kinase 1/SPHK1 in Human Prostate. Sphingosine Kinase 1/SPHK1 was detected in immersion fixed paraffin-embedded sections of human prostate using Sheep Anti-Human Sphingosine Kinase 1/SPHK1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5536) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Human Sphingosine Kinase 1/SPHK1 ELISA Standard Curve. Recombinant Human Sphingosine Kinase 1/SPHK1 protein was serially diluted 2-fold and captured by Mouse Anti-Human Sphingosine Kinase 1/SPHK1 Monoclonal Antibody (Catalog # MAB5536) coated on a Clear Polystyrene Microplate (Catalog # DY990). Sheep Anti-Human Sphingosine Kinase 1/SPHK1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5536) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Sphingosine Kinase 1/SPHK1
Sphingosine kinases are cytosolic or membrane-associated enzymes that catalyze the phosphorylation of sphingosine to sphingosine-1-phosphate (S1P). Two types of sphingosine kinases, SPHK1 and SPHK2, are known to be expressed in human cells. The two enzymes share considerable amino acid sequence similarity, but differ in their N‑terminal and central regions (1). The two proteins also differ in tissue distribution and some kinetic properties (1). S1P is a lipid messenger that regulates diverse physiological processes including cell proliferation, migration, apoptosis, inflammation, calcium homeostasis and cytoskeletal structure (2, 3). The level of S1P is tightly controlled by SPHKs and S1P degrading enzymes. SPHK1 and its activation can be stimulated by several growth factors such as tumor necrosis factor-alpha, epidermal growth factor and transforming growth factor-beta (3, 4). Expression of SPHK1 has been found to increase in many human solid tumors and overexpression of SPHK1 is associated with tumor angiogenesis (5). Such studies have implicated SPHK1 as a new target for cancer treatment.
- Liu, H. et al. (2000) J. Biol. Chem. 275:19513.
- Spiegel, S. (1999) J. Leukocyte Biol. 65:341.
- Alemany, R. et al. (2007) Naunyn-Schmiedegerg’s Arch. Pharmacol. 374:413.
- Pederson, L. et al. (2008) Proc. Natl. Acad. Scis USA. 105:20764.
- Shida, D. et al. (2008) Curr. Drug Targets. 9:662.
Citation for Human Sphingosine Kinase 1/SPHK1 Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
1 Citation: Showing 1 - 1
Pseudomonas-derived ceramidase induces production of inflammatory mediators from human keratinocytes via sphingosine-1-phosphate.
Authors: Oizumi A, Nakayama H, Okino N, Iwahara C, Kina K, Matsumoto R, Ogawa H, Takamori K, Ito M, Suga Y, Iwabuchi K
PLoS ONE, 2014;9(2):e89402.
Sample Types: Cell Lysates
Applications: Western Blot
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Cell and Tissue Staining Kits
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