S1PR1 antibody (C-Term)

Catalog No. ABIN2857018

This independently validated Rabbit Polyclonal antibody specifically detects S1PR1 in WB, IF, IHC (p) and ICC. It exhibits reactivity toward Human.

Quick Overview for S1PR1 antibody (C-Term) (ABIN2857018)

Target: S1PR1 (Sphingosine-1-Phosphate Receptor 1 (S1PR1))

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This S1PR1 antibody is un-conjugated

Application: Western Blotting (WB), Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunocytochemistry (ICC)

Product Details anti-S1PR1 Antibody

Binding Specificity: C-Term

Cross-Reactivity: Human, Mouse

Characteristics: Rabbit Polyclonal antibody to EDG1 (sphingosine-1-phosphate receptor 1)
EDG1 antibody [C2C3], C-term

Purification: Purified by antigen-affinity chromatography.

Immunogen: Carrier-protein conjugated synthetic peptide encompassing a sequence within the C-terminus region of human EDG1. The exact sequence is proprietary.

Isotype: IgG

Application Details

Application Notes: WB: 1:500-1:3000. ICC/IF: 1:100-1:1000. IHC-P: 1:100-1:1000. Optimal dilutions/concentrations should be determined by the researcher. Not tested in other applications.

Comment:

Positive Control: HepG2

Restrictions: For Research Use only

Independent Validation (IF)

Validation #100014 (Immunofluorescence)

by: Memorial Sloan-Kettering Cancer Center, Parada lab

No.: #100014

Date: 05/11/2016

Antigen: Sphingosine-1-Phosphate Receptor 1 (S1PR1)

Lot Number: 39471

Method validated: Immunofluorescence

Positive Control: Wild-type mouse adult brain cryo-sections

Negative Control: Rabbit IgG

Notes: The antibody showed signals along the subventricular zone, which is consistent with the RNA expression analysis.

Validation Images

IF validation of S1PR1 antibody ABIN2857018 on mouse brain sections. A. ABIN2857018 diluted 1:200 (red), DAPI counterstaining (blue). B. Dilution factor 1:100. C. Dilution factor 1:200. D. Dilution factor 1:400.

Full Methods

Primary Antibody: ABIN2857018

Secondary Antibody: Donkey-anti-Rabbit-Cy3

Full Protocol:

• Wild-type mouse adult brain was dissected, fixed in 4% PFA for over/night, saturated in 30% sucrose, mounted in Cryo, and sectioned into 12µm sections.
• Immunostaining for Cryo Sections:
• • Dry slides at 50-55°C for 15-30min (or RT for 30min).
  • Wash with PBS 5min X 3 (shake gently).
  • Block with 10% Normal Donkey Serum (NDS) at RT for 1h.
  • Incubate with ABIN2857018 diluted 1:10 - 1:800 in 2% NDS at 4°C overnight.
  • Wash with PBS 5min X 3.
  • Incubate with the secondary antibody Donkey-anti-Rabbit-Cy3 together with DAPI 1:100 at RT for 1h (prepared in 2% NDS), cover the chamber with foil.
  • Wash with PBS 5min X 3.
  • Add three drops of water-based Immu-mount.
  • Cover with 24 X 60 cover glass.
• Imaging:
• • Images were taken using a Zeiss Axio2-Imaging system, 20X-0.5NA-Dry Dye-1004-072 objective, DAPI and Cy3 channel.

Experimental Notes: ABIN2857018 worked the best at 1:200. At 1:400, or 1:800, it seems to lose some signals in some cells.

Handling

Format: Liquid

Concentration: 1 mg/mL

Buffer: 1XPBS ( pH 7), 20 % Glycerol, 0.01 % Thimerosal

Preservative: Thimerosal (Merthiolate)

Precaution of Use: This product contains Thimerosal (Merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4°C. For long-term storage, aliquot and store at -20°C or below. Avoid multiple freeze-thaw cycles.

Target Details for S1PR1

Target: S1PR1 (Sphingosine-1-Phosphate Receptor 1 (S1PR1))

Alternative Name: sphingosine-1-phosphate receptor 1

Background: The protein encoded by this gene is structurally similar to G protein-coupled receptors and is highly expressed in endothelial cells. It binds the ligand sphingosine-1-phosphate with high affinity and high specificity, and suggested to be involved in the processes that regulate the differentiation of endothelial cells. Activation of this receptor induces cell-cell adhesion.

Cellular Localization: Cell membrane, Multi-pass membrane protein

Molecular Weight: 43 kDa

Gene ID: 1901

UniProt: P21453

Pathways: Signaling Events mediated by VEGFR1 and VEGFR2