Revision 3
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-IC

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

40-50

Source/Isotype:

Rabbit IgG

UniProt ID:

#P21453

Entrez-Gene Id:

1901

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:1600 - 1:3200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Sphingosine-1-Phosphate Receptor 1 (E8U3O) Rabbit mAb recognizes endogenous levels of total sphingosine-1-phosphate receptor 1 protein. Nonspecific nuclear background may be observed by immunofluorescence in some cell lines, especially with higher than recommended concentrations of antibody.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly366 of human sphingosine-1-phosphate receptor 1 protein.

Background

Sphingosine-1-phosphate receptor 1 (S1PR1, EDG1) is a G protein-coupled receptor (GPCR), one of five members (S1PR1-5) belonging to the sphingosine-1-phosphate receptor (S1PR) family (1,2). This family of GPCRs has been shown to exhibit broad and often overlapping expression patterns across a wide array of tissues. The cognate ligand for S1PRs is sphingosine-1-phosphate (S1P), a phosphorylated lipid signaling molecule derived from sphingosine. S1P/S1PR binding can trigger the activation and/or inhibition of various downstream signaling pathways, depending on context. Pathways identified downstream of S1P/S1PR include PI3K/Akt, PLC, Ras/MAPK and PKA (3,4). S1P/S1PR1 signaling pathways play important roles in many basic cellular functions during normal development, including chemotaxis, vascular integrity, and immune cell egress, but can also contribute to a variety of pathological conditions, including atherogenesis, multiple sclerosis, and tumorigenesis (5-9).

  1. Pérez-Jeldres, T. et al. (2021) Drugs 81, 985-1002.
  2. Chun, J. et al. (2021) Drugs 81, 207-231.
  3. Pan, Y. et al. (2021) Clin Exp Pharmacol Physiol 48, 637-650.
  4. Xiao, L. et al. (2019) Front Immunol 10, 1409.
  5. Rivera, J. et al. (2008) Nat Rev Immunol 8, 753-63.
  6. Anwar, M. and Mehta, D. (2020) Biochim Biophys Acta Mol Cell Biol Lipids 1865, 158760.
  7. Song, J.H. et al. (2021) Biomol Ther (Seoul) 29, 373-383.
  8. Roy, R. et al. (2021) CNS Drugs 35, 385-402.
  9. Anu, B. et al. (2021) Adv Protein Chem Struct Biol 125, 259-274.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 3
#63335

Sphingosine-1-Phosphate Receptor 1 (E8U3O) Rabbit mAb

Western Blotting Image 1: Sphingosine-1-Phosphate Receptor 1 (E8U3O) Rabbit mAb Expand Image
Western blot analysis of extracts from SK-HEP-1 and DU 145 cells using Sphingosine-1-Phosphate Receptor 1 (E8U3O) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The absence of sphingosine-1-phosphate receptor 1 protein in DU 145 cells is consistent with mRNA expression profiles reported in public bioinformatic databases, confirming specificity of the antibody for sphingosine-1-phosphate receptor 1.
Western Blotting Image 2: Sphingosine-1-Phosphate Receptor 1 (E8U3O) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines and tissues using Sphingosine-1-Phosphate Receptor 1 (E8U3O) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The absence of sphingosine-1-phosphate receptor 1 protein in DU 145 cells is consistent with mRNA expression profiles reported in public bioinformatic databases, confirming specificity of the antibody for sphingosine-1-phosphate receptor 1.
Immunoprecipitation Image 1: Sphingosine-1-Phosphate Receptor 1 (E8U3O) Rabbit mAb Expand Image
Immunoprecipitation of sphingosine-1-phosphate receptor 1 protein from Sup-M2 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Sphingosine-1-Phosphate Receptor 1 (E8U3O) Rabbit mAb. Western blot analysis was performed using Sphingosine-1-Phosphate Receptor 1 (E8U3O) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Immunofluorescence Image 1: Sphingosine-1-Phosphate Receptor 1 (E8U3O) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of SK-HEP-1 cells (left, positive), OVCAR-8 cells (middle, positive), and DU 145 cells (right, negative) using Sphingosine-1-Phosphate Receptor 1 (E8U3O) Rabbit mAb (green), COX IV (4D11-B3-E8) Mouse mAb #11967 (red), and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).