Name: Anti-S1PR3 (EDG3) (extracellular)-ATTO Fluor-488 Antibody
Brand: Alomone Labs
SKU: ASR-013-AG

Overview

Cat #: ASR-013-AG

Alternative Name Sphingosine 1-phosphate receptor 3, S1P3, Endothelial differentiation G-protein coupled receptor 3, Lysophospholipid receptor B3, LPB3

Lyophilized Powder yes

Type: Polyclonal

Host: Rabbit

Reactivity: h, m, r

Immunogen

Peptide (C)DYVGKLAGRLRD, corresponding to amino acid residues 23-34 of mouse S1PR3 (Accession Q9Z0U9). Extracellular, N-terminus.

Accession (Uniprot) Number Q9Z0U9

Gene ID 13610

Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.

Homology Rat - identical; human - 9/12 amino acid residues identical.

RRID AB_10919249.

Purity Affinity purified on immobilized antigen.

Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN₃.

Isotype Rabbit IgG.

Label ATTO-488. Maximum absorption 501 nm; maximum fluorescence 523 nm. The fluorescence is excited most efficiently in the 480 – 515 nm range. This label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC.

Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.

Reconstitution 50 µl double distilled water (DDW).

Antibody concentration after reconstitution 1 mg/ml.

Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 × g 5 min).

Standard quality control of each lot Western blot analysis (unlabeled antibody, #ASR-013), and direct flow cytometry (labeled antibody).

Applications: fc

May also work in: ic*, ih*, lci*

Direct flow cytometry

Cell surface detection of S1PR3 in human Jurkat intact live cells:
^___ Unstained cells.
^___ Cells + Anti-S1PR3 (EDG3) (extracellular)-ATTO Fluor-488 Antibody (#ASR-013-AG), (5-10 µg/5x10⁵ cells).

References

Hla, T. and Brinkmann, V. (2011) Neurology 76, S3.
Rosen, H. et al. (2007) Trends Immunol. 28, 102.
Rosen, H. et al. (2009) Annu. Rev. Biochem. 78, 743.
Jiang, L.I. et al. (2007) J. Biol. Chem. 282, 10576.
Sanchez, T. and Hla, T. (2004) J. Cell. Biochem. 92, 913.

Scientific background

Sphingosine 1-phosphate (S1P) is an active byproduct of sphingomyelin metabolism. All cells can synthesize this biomolecule but the majority of its synthesis comes from erythrocytes and endothelial cells. In cases of inflammation, mast cells and platelets are the main source of sphingosine 1-phosphate¹. S1P cellular functions could be intracellular, where it is synthesized. In addition, upon secretion, it circulates in the blood via its binding to high-density lipoproteins and albumin. When S1P reaches its target cell it could activate 5 different high affinity receptors belonging to the G-protein coupled receptor superfamily: Sphingosine 1-phosphate receptors, termed S1PR1-5¹.

Stimulation of S1P receptors triggers a cascade of signaling events depending on the receptor and on the G-protein it couples. S1PR1 couples to G_i². S1PR2 can couple to G_s, G_q or G_12/13 and S1PR3-5 can couple to G_ior G_12/13^3-4. The pathways activated vary from Ca²⁺ mobilization, activation or inhibition of adenylate cyclase phospholipase C activation and more. Through the different signaling pathways these receptors activate, S1P1 receptors are implicated in adherens junction assembly, cytoskeletal changes, cell migration, proliferation and apoptosis⁵.

S1PR3 is specifically detected in the brain, heart, spleen, liver, lung, thymus, kidney, testis and skeletal muscle³. S1PR1 and S1PR2 are generally expressed in the CNS, cardiovascular and immune systems¹. S1PR4 is specifically expressed in the lymphoid tissue and S1PR5 in natural killer cells and olygodendrocytes¹.

Malfunction of S1P receptor signaling is reported in various disorders, for example multiple sclerosis and may be targets for the development of therapeutic drugs¹.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat

Lyophilized Powder

Anti-S1PR3 (EDG3) (extracellular) Antibody (#ASR-013) is a highly specific antibody directed against an extracellular epitope of mouse sphingosine 1-phosphate receptor 3. The antibody can be used in western blot and indirect flow cytometry applications. It has been designed to recognize S1PR3 from human, rat and mouse samples.

Anti-S1PR3 (EDG3) (extracellular)-ATTO Fluor-488 Antibody (#ASR-013-AG) is directly labeled with an ATTO-488 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-488 label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC. Anti-S1PR3 (EDG3) (extracellular)-ATTO Fluor-488 Antibody is especially suited for experiments requiring simultaneous labeling of different markers.

Certificate of Analysis SDS

For research purposes only, not for human use

Last Update: 02/01/2024

Applications

Citations

Specifications

Scientific Background

Specific Control Products

S1PR3/EDG3 (extracellular) Blocking Peptide (#BLP-SR013) is the original antigen used for immunization during Anti-S1PR3 (EDG3) (extracellular) Antibody (#ASR-013) generation. The blocking peptide binds and ‘blocks’ Anti-S1PR3/EDG3 (extracellular) primary antibody, this makes it a good negative reagent control to help confirm antibody specificity in western blot and immunohistochemistry applications. This control is also often called a pre-adsorption control.

S1PR3/EDG3 (extracellular) Blocking Peptide (#BLP-SR013)
Isotype controls are primary antibodies that lack specificity to the target, but match the class and type of the primary antibody used in the application. Isotype controls are used as negative controls to help differentiate non-specific background signal from specific antibody signal.
Our Rabbit IgG Isotype Control-ATTO Fluor-488 (#RIC-001-AG), was specifically developed to be used as a negative control in immunohistochemistry and surface staining flow cytometry applications, along our line of rabbit primary antibodies conjugated to the ATTO Fluor-488 fluorophore.

Rabbit IgG Isotype Control-ATTO Fluor-488 (#RIC-001-AG)

Shipping and Ordering Information

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Anti-S1PR3 (EDG3) (extracellular)-ATTO Fluor-488 Antibody