Overview

  • Product name

  • Description

    Rabbit polyclonal to EDG8
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Dog, Pig
  • Immunogen

    Synthetic peptide corresponding to Human EDG8 aa 1-100 conjugated to keyhole limpet haemocyanin.
    Database link: Q91X56
    (Peptide available as ab103906)

  • Positive control

    • This antibody gave a positive signal in the following lysates: Brain: corpus callosum (Human) Whole Cell Lysate; Brain (Human) Tissue Lysate; Spleen (Human) Tissue Lysate; Lung (Human) Tissue Lysate; U937 (Human leukemic monocyte lymphoma cell line) Whole Cell Lysate; MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab92994 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 10 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use at an assay dependent concentration. Detects a band of approximately 41 kDa (predicted molecular weight: 42 kDa).

Target

  • Function

    Receptor for the lysosphingolipid sphingosine 1-phosphate (S1P). S1P is a bioactive lysophospholipid that elicits diverse physiological effect on most types of cells and tissues. Is coupled to both the G(i/0)alpha and G(12) subclass of heteromeric G-proteins (By similarity). May play a regulatory role in the transformation of radial glial cells into astrocytes and may affect proliferative activity of these cells.
  • Tissue specificity

    Widely expressed in the brain, most prominently in the corpus callosum, which is predominantly white matter. Detected in spleen, peripheral blood leukocytes, placenta, lung, aorta and fetal spleen. Low-level signal detected in many tissue extracts. Overexpressed in leukemic large granular lymphocytes. Isoform 1 is predominantly expressed in peripheral tissues. Isoform 2 is expressed in brain, spleen and peripheral blood leukocytes.
  • Sequence similarities

    Belongs to the G-protein coupled receptor 1 family.
  • Developmental stage

    At 24 weeks of gestation, fragments of radial glial fibers are positive within the cortical plate and subplate of allocortical areas. These positive fragments often appear enlarged as varicosities and some of them terminate at blood vessels. Between 28 and 30 weeks of gestation, all iso- and allocortical areas contain immunolabelled radial glial fibers revealing curvature next to sulci. After 32 weeks of gestation, radial glial fibers gradually disappear; instead positive transitional stages between radial glia and astrocytes were found.
  • Cellular localization

    Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • EDG 8 antibody
    • EDG8 S1P receptor antibody
    • Endothelial differentiation G protein coupled receptor 8 antibody
    • Endothelial differentiation G-protein-coupled receptor 8 antibody
    • Endothelial Differentiation Gene 8 antibody
    • Endothelial differentiation sphingolipid G protein coupled receptor 8 antibody
    • S1P receptor 5 antibody
    • S1P receptor Edg 8 antibody
    • S1P receptor Edg-8 antibody
    • S1P receptor Edg8 antibody
    • S1P5 antibody
    • S1PR5 antibody
    • S1PR5_HUMAN antibody
    • Sphingosine 1 Phosphate Receptor 5 antibody
    • Sphingosine 1 phosphate receptor Edg 8 antibody
    • Sphingosine 1 phosphate receptor Edg8 antibody
    • Sphingosine 1-phosphate receptor 5 antibody
    • Sphingosine 1-phosphate receptor Edg-8 antibody
    • SPPR 1 antibody
    • SPPR 2 antibody
    • SPPR1 antibody
    • SPPR2 antibody
    see all

Images

  • All lanes : Anti-EDG8 antibody (ab92994) at 1 µg/ml

    Lane 1 : Human brain corpus callosum tissue lysate (alzheimer) - total protein (ab30027)
    Lane 2 : Human brain tissue lysate - total protein (ab29466)
    Lane 3 : Human spleen tissue lysate - total protein (ab29699)
    Lane 4 : Lung (Human) Tissue Lysate
    Lane 5 : U937 (Human leukemic monocyte lymphoma cell line) Whole Cell Lysate
    Lane 6 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 41 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 30 kDa, 80 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 8 minutes
  • ICC/IF image of ab92994 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab92994 at 10µg/ml overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti- rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • IHC image of EDG8 staining in human hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab92994, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

References

This product has been referenced in:

  • Palangi A  et al. Differential expression of S1P receptor subtypes in human bladder transitional cell carcinoma. Clin Transl Oncol N/A:N/A (2019). Read more (PubMed: 30712233) »
  • Arana L  et al. Ceramide 1-phosphate induces macrophage chemoattractant protein-1 release: involvement in ceramide 1-phosphate-stimulated cell migration. Am J Physiol Endocrinol Metab 304:E1213-26 (2013). Read more (PubMed: 23548612) »
See all 2 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Abreviews
Application
Western blot
Sample
Mouse Tissue lysate - other (Synaptosomes Cortex)
Gel Running Conditions
Reduced Denaturing (Bis-Tris 4-12% gel)
Loading amount
15 µg
Specification
Synaptosomes Cortex
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted May 08 2019

Question
Answer

Thank you for contacting us. I am very sorry for inconvenience caused due to un-publishing of ab13130. We have other EDG8 antibodies ab92994, ab60039 and ab89677 available in catalogue.As we will not be able to provide the free of charge samples of these products so we would like to familiarize you with the Abreview discount programme. Please click to below link to read the terms and conditions; https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount As discussed I am also happy in providing XXXXX discount on the purchase of ab92994. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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