Overview

  • Product name

    Anti-ENPP2/ATX antibody [5H3] - BSA and Azide free
    See all ENPP2/ATX primary antibodies
  • Description

    Mouse monoclonal [5H3] to ENPP2/ATX - BSA and Azide free
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, ELISA, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant full length protein corresponding to Human ENPP2/ATX.

  • Positive control

    • WB: HEK293 whole cell lysate and in the following human tissue lysates: kidney; placenta; ovary; small intestine. IHC: human tonsil paraffin sections. ICC/IF: HeLa cells.
  • General notes

    ab238670 is a PBS only version of ab77104.

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Applications

Our Abpromise guarantee covers the use of ab238670 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 99 kDa).
ELISA Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml.
ICC/IF Use a concentration of 10 µg/ml.

Target

  • Function

    Hydrolyzes lysophospholipids to produce lysophosphatidic acid (LPA) in extracellular fluids. Major substrate is lysophosphatidylcholine. Also can act on sphingosylphosphphorylcholine producing sphingosine-1-phosphate, a modulator of cell motility. Can hydrolyze, in vitro, bis-pNPP, to some extent pNP-TMP, and barely ATP. Involved in several motility-related processes such as angiogenesis and neurite outgrowth. Acts as an angiogenic factor by stimulating migration of smooth muscle cells and microtubule formation. Stimulates migration of melanoma cells, probably via a pertussis toxin-sensitive G protein. May have a role in induction of parturition. Possible involvement in cell proliferation and adipose tissue development. Tumor cell motility-stimulating factor.
  • Tissue specificity

    Predominantly expressed in brain, placenta, ovary, and small intestine. Expressed in a number of carcinomas such as hepatocellular and prostate carcinoma, neuroblastoma and non-small-cell lung cancer. Expressed in body fluids such as plasma, cerebral spinal fluid (CSF), saliva, follicular and amniotic fluids. Not detected in leukocytes. Isoform 1 is more highly expressed in peripheral tissues than in the central nervous system (CNS). Adipocytes only express isoform 1. Isoform 3 is more highly expressed in the brain than in peripheral tissues.
  • Sequence similarities

    Belongs to the nucleotide pyrophosphatase/phosphodiesterase family.
    Contains 2 SMB (somatomedin-B) domains.
  • Post-translational
    modifications

    N-glycosylation, but not furin-cleavage, plays a critical role on secretion and on lysoPLD activity.
  • Cellular localization

    Secreted. Secreted by most body fluids including serum and CSF. Also by adipocytes and numerous cancer cells.
  • Information by UniProt
  • Database links

  • Alternative names

    • ATX antibody
    • ATX X antibody
    • Autotaxin antibody
    • Autotaxin t antibody
    • E NPP 2 antibody
    • E-NPP 2 antibody
    • Ectonucleotide pyrophosphatase/phosphodiesterase 2 antibody
    • Ectonucleotide pyrophosphatase/phosphodiesterase family member 2 antibody
    • Enpp2 antibody
    • ENPP2_HUMAN antibody
    • Extracellular lysophospholipase D antibody
    • FLJ26803 antibody
    • LysoPLD antibody
    • NPP2 antibody
    • PD IALPHA antibody
    • PDNP2 antibody
    • Phosphodiesterase I alpha antibody
    • Phosphodiesterase I/nucleotide pyrophosphatase 2 antibody
    • Plasma lysophospholipase D antibody
    see all

Images

  • All lanes : Anti-ENPP2/ATX antibody [5H3] (ab77104) at 10 µg/ml

    Lane 1 : ab30203
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 99 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 180 kDa, 250 kDa, 50 kDa, 65 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes


    This data was developed using the same antibody clone in a different formulation containing PBS, Azide and arginine (ab77104).

  • IHC image of ENPP2/ATX staining in Human Tonsil FFPE section, performed on a 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 ab77104, 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.

    This data was developed using the same antibody clone in a different formulation containing PBS, Azide and arginine (ab77104).

  • ICC/IF image of ab77104 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab77104, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

    This data was developed using the same antibody clone in a different formulation containing PBS, Azide and arginine (ab77104).

  • All lanes : Anti-ENPP2/ATX antibody [5H3] (ab77104) at 10 µg/ml

    Lane 1 : ab29745
    Lane 2 : ab30222
    Lane 3 : ab29276

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Predicted band size: 99 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?
    Additional bands at: 300 kDa, 40 kDa, 55 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes


    ENPP2/ATX contains three potential glycosylation sites (SwissProt), which might explain its migration at a higher molecular weight than predicted.

    This data was developed using the same antibody clone in a different formulation containing PBS, Azide and arginine (ab77104).

References

ab238670 has not yet been referenced specifically in any publications.

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