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    enpp2atx-antibody-ab140915.pdf

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Anti-ENPP2/ATX antibody (ab140915)

  • Datasheet
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Western blot - Anti-ENPP2/ATX antibody (ab140915)
  • Immunocytochemistry/ Immunofluorescence - Anti-ENPP2/ATX antibody (ab140915)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENPP2/ATX antibody (ab140915)

Key features and details

  • Rabbit polyclonal to ENPP2/ATX
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Human
  • Isotype: IgG

You may also be interested in

Protein
Product image
Recombinant Human ENPP2/ATX protein (ab126918)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Knockout
Product image
Human ENPP2 (ATX) knockout HeLa cell line (ab265377)

View more associated products

Overview

  • Product name

    Anti-ENPP2/ATX antibody
    See all ENPP2/ATX primary antibodies
  • Description

    Rabbit polyclonal to ENPP2/ATX
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human ENPP2/ATX aa 300-400 conjugated to keyhole limpet haemocyanin.
    Database link: Q13822

  • Positive control

    • WB: MOLT4, HeLa, HepG2 and Y79 whole cell lysates. IHC-P: FFPE human tonsil tissue sections. ICC/IF: U-87 MG cells.
  • General notes

     This product was previously labelled as ENPP2

     

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipid Metabolism
    • Lipases
    • Cardiovascular
    • Angiogenesis
    • Angiogenic Factors
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Other
    • Signal Transduction
    • Metabolism
    • Lipid metabolism
    • Cancer
    • Tumor biomarkers
    • Enzymes
    • Other
    • Cardiovascular
    • Atherosclerosis
    • Thrombosis
    • Other
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipases
    • Metabolism
    • Types of disease
    • Heart disease

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human ENPP2/ATX protein (ab126918)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab140915 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IHC-P
Human
WB
Human
All applications
Mouse
Rat
Rabbit
Horse
Cow
Dog
Pig
Chimpanzee
Macaque monkey
Gorilla
Orangutan
Application Abreviews Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 125 kDa (predicted molecular weight: 98 kDa).
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
Use a concentration of 10 µg/ml.
Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 125 kDa (predicted molecular weight: 98 kDa).
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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.
  • Target information above from: UniProt accession Q13822 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 532663 Cow
    • Entrez Gene: 5168 Human
    • Entrez Gene: 18606 Mouse
    • Entrez Gene: 84050 Rat
    • Omim: 601060 Human
    • SwissProt: A1A4K5 Cow
    • SwissProt: Q13822 Human
    • SwissProt: Q9R1E6 Mouse
    • SwissProt: Q64610 Rat
    • Unigene: 190977 Human
    • Unigene: 250256 Mouse
    • Unigene: 20403 Rat
    see all
  • 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

  • Western blot - Anti-ENPP2/ATX antibody (ab140915)
    Western blot - Anti-ENPP2/ATX antibody (ab140915)
    All lanes : Anti-ENPP2/ATX antibody (ab140915) at 1 µg/ml

    Lane 1 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
    Lane 5 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate with Immunising peptide at 1 µg/ml
    Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Immunising peptide at 1 µg/ml
    Lane 7 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate with Immunising peptide at 1 µg/ml
    Lane 8 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate with Immunising peptide at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 98 kDa
    Observed band size: 125 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 21 kDa (possible non-specific binding), 32 kDa (possible non-specific binding)


    Exposure time: 4 minutes


    ENPP2/ATX contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab140915 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

  • Immunocytochemistry/ Immunofluorescence - Anti-ENPP2/ATX antibody (ab140915)
    Immunocytochemistry/ Immunofluorescence - Anti-ENPP2/ATX antibody (ab140915)

    ICC/IF image of ab140915 stained U-87 MG 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 (ab140915, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENPP2/ATX antibody (ab140915)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENPP2/ATX antibody (ab140915)

    IHC image of ENPP2/ATX staining in human tonsil formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab140915, 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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (2)

    Publishing research using ab140915? Please let us know so that we can cite the reference in this datasheet.

    ab140915 has been referenced in 2 publications.

    • Cha YJ & Koo JS Expression of Autotaxin?Lysophosphatidate Signaling-Related Proteins in Breast Cancer with Adipose Stroma. Int J Mol Sci 20:N/A (2019). PubMed: 31035435
    • Shim SJ  et al. The expressions of autotaxin-lysophosphatidate signaling-related proteins in metastatic breast cancer. Int J Clin Exp Pathol 12:2920-2930 (2019). PubMed: 31934128

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