Human Sphingosine Kinase 1/SPHK1 Antibody

(1 citations)   
  • Species Reactivity
    Human
  • 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
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human Sphingosine Kinase 1/SPHK1
    Asp2-Leu398
    Accession # NP_068807
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunoprecipitation
    25 µg/mL
    Conditioned cell culture medium spiked with Recombinant Human SPHK1 (Catalog #
    5536-SK), see our available Western blot detection antibodies
  • Immunocytochemistry
    5-15 µg/mL
    See below
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.
Data Examples
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.
Immunocytochemistry
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.
Immunohistochemistry
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.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.2 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & 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.
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.
  • References:
    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.
  • Entrez Gene IDs:
    8877 (Human); 20698 (Mouse); 170897 (Rat)
  • Alternate Names:
    EC 2.7.1.91; SK1; sphingosine kinase 1; SPHK1; SPHKSK 1; SPK 1; SPK; SPK1
Related Research Areas
Citations:

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 Citations: Showing 1 - 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.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
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Immunohistochemistry Reagents
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Mounting Medium

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Antigen Retrieval Reagent Sampler (50 mL each)

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Antigen Retrieval Reagent-Acidic

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Isotype Controls
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Normal Sheep IgG Control

Ctrl 5-001-A 9
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Secondary Antibodies
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Sheep IgG HRP-conjugated Antibody

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Donkey Anti-Sheep IgG NorthernLights™ NL557-conjugated Antibody

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Donkey Anti-Sheep IgG NorthernLights™ NL637-conjugated Antibody

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Donkey Anti-Sheep IgG NorthernLights™ NL493-conjugated Antibody

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Sheep IgG (H+L) PE-conjugated Antibody

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Donkey Anti-Sheep IgG Biotinylated Antibody

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Sheep IgG VisUCyte HRP Polymer Antibody

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Sheep IgG (H+L) APC-conjugated Antibody

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Donkey Anti-Sheep IgG (H+L) Fluorescein-conjugated Antibody

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Donkey Anti-Sheep IgG (H+L) PerCP-conjugated Antibody

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