Human Sphingosine Kinase 1/SPHK1 Antibody

Catalog # Availability Size / Price Qty
AF5536
AF5536-SP
Human Sphingosine Kinase 1/SPHK1 Antibody in Data
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Human Sphingosine Kinase 1/SPHK1 Antibody Summary

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
ELISA

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.

 
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

Western Blot Detection of Human Sphingosine Kinase 1/SPHK1 antibody by Western Blot. View Larger

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 antibody in HepG2 Human Cell Line by Immunocytochemistry (ICC). View Larger

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 antibody in Human Prostate by Immunohistochemistry (IHC-P). View Larger

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).

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

Product Datasheets

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

  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 Types: Cell Lysates
    Applications: Western Blot

FAQs

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Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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