Sphingosine Kinase 1/SPHK1 Antibody [Unconjugated]

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 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 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.

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).

• Reactivity: Hu
• Applications: WB, ELISA, IHC, IP, ICC/IF
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Sphingosine Kinase 1/SPHK1 Antibody [Unconjugated] Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf 21-derived recombinant human Sphingosine Kinase 1/SPHK1
  Asp2-Leu398
  Accession # NP_068807
• Specificity: Detects human SPHK1 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human SPHK2 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: SPHK1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Immunoprecipitation 25 ug/mL
  • Western Blot 0.5 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Sphingosine Kinase 1/SPHK1 Antibody [Unconjugated]

• EC 2.7.1.91
• SK1
• sphingosine kinase 1
• SPHK1
• SPHKSK 1
• SPK 1
• SPK
• SPK1

Background

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.

1. Liu, H. et al. (2000) J. Biol. Chem. 275:19513.
2. Spiegel, S. (1999) J. Leukocyte Biol. 65:341.
3. Alemany, R. et al. (2007) Naunyn-Schmiedegerg’s Arch. Pharmacol. 374:413.
4. Pederson, L. et al. (2008) Proc. Natl. Acad. Scis USA. 105:20764.
5. Shida, D. et al. (2008) Curr. Drug Targets. 9:662.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Sphingosine Kinase 1/SPHK1 Antibody (AF5536)(2)

Publications using AF5536

• G Li, H Xie, X Cao, C Ma, Y Li, L Chen Ginsenoside Rg1 exerts anti?apoptotic effects on non?alcoholic fatty liver cells by downregulating the expression of SGPL1 Molecular Medicine Reports, 2022-03-24;25(5):. 2022-03-24 [PMID: 35322862] (Western Blot, Human)
• Oizumi A, Nakayama H, Okino N, Iwahara C, Kina K, Matsumoto R, Ogawa H, Takamori K, Ito M, Suga Y, Iwabuchi K Pseudomonas-derived ceramidase induces production of inflammatory mediators from human keratinocytes via sphingosine-1-phosphate. PLoS ONE, 2014-02-25;9(2):e89402. 2014-02-25 [PMID: 24586752] (Western Blot, Human)

Bioinformatics

• Gene Symbol: SPHK1
• Uniprot:
  • NP_068807()