EDG1 Recombinant Rabbit Monoclonal Antibody [JM10-66]
cat.: ET1703-27
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: JM10-66
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 43 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human EDG1 aa 2-51 / 382.
Positive control: SH-SY5Y cell lysate, HepG2 cell lysate, bEnd.3 cell lysate, HepG2, HUVEC, SH-SY5Y, human breast carcinoma tissue, human liver tissue, mouse brain tissue, mouse heart tissue, Jurkat.
Subcellular location: Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:2,000-5,000
1:100-1:500
1:100-1:500
1:50-1:200
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: P21453 Human | O08530 Mouse | P48303 Rat
Alternative names: CD363 CHEDG 1 CHEDG1 D1S3362 ECGF 1 ECGF1 EDG 1 EDG1 endothelial differentiation G protein coupled receptor 1 Endothelial differentiation G-protein coupled receptor 1 Endothelial differentiation sphingolipid G protein coupled receptor 1 FLJ58121 G protein coupled sphingolipid receptor g protein-coupled receptor edg-1 S1P receptor 1 S1P receptor Edg 1 S1P receptor Edg-1 S1P receptor Edg1 S1P(1) receptor S1P1 s1pr1 S1PR1_HUMAN sphingolipid g-protein-coupled receptor 1 Sphingosine 1 phosphate receptor Edg 1 Sphingosine 1 phosphate receptor EDG1 sphingosine 1- phosphate receptor 1 Sphingosine 1-phosphate receptor 1 Sphingosine 1-phosphate receptor Edg-1 Sphingosine 1phosphate receptor type 1 S1P1
Images
ET1703-27_1.jpg Fig1: Western blot analysis of EDG1 on different lysates with Rabbit anti-EDG1 antibody (ET1703-27) at 1/5,000 dilution.

Lane 1: SH-SY5Y cell lysate
Lane 2: HepG2 cell lysate
Lane 3: bEnd.3 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 43 kDa
Observed band size: 43 kDa

Exposure time: 1 minute 21 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-27) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.
ET1703-27_2.jpg Fig2: Western blot analysis of EDG1 on different lysates with Rabbit anti-EDG1 antibody (ET1703-27) at 1/5,000 dilution.

Lane 1: HepG2-si NT cell lysate
Lane 2: HepG2-si EDG1 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 43 kDa
Observed band size: 43 kDa

Exposure time: 2 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-27) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1703-27_3.jpg Fig3: Western blot analysis of EDG1 on different lysates with Rabbit anti-EDG1 antibody (ET1703-27) at 1/2,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: PC-12 cell lysate
Lane 3: Rat brain tissue lysate
Lane 4: Rat spleen tissue lysate

Lysates/proteins at 20/40 µg/Lane.

Predicted band size: 43 kDa
Observed band size: 43 kDa

Exposure time: 60 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-27) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1703-27_4.jpg Fig4: ICC staining of EDG1 in HepG2 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-27, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-27_5.jpg Fig5: ICC staining of EDG1 in HUVEC cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-27, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-27_6.jpg Fig6: ICC staining of EDG1 in SH-SY5Y cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-27, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-27_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-EDG1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-27, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-27_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-EDG1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-27, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-27_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-EDG1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-27, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-27_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-EDG1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-27, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-27_11.jpg Fig11: Flow cytometric analysis of EDG1 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1703-27, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1703-27_12.jpg Fig12: EDG1 was immunoprecipitated in 0.2mg HepG2 cell lysate with ET1703-27 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1703-27 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HepG2 cell lysate (input)
Lane 2: ET1703-27 IP in HepG2 cell lysate
Lane 3: Rabbit IgG instead of ET1703-27 in HepG2 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 4 mintues
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.