Product nameAnti-ENPP2/ATX antibody
See all ENPP2/ATX primary antibodies
DescriptionRabbit polyclonal to ENPP2/ATX
Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat, Rabbit, Horse, Cow, Dog, Pig, Chimpanzee, Macaque monkey, Gorilla, Orangutan
Synthetic peptide corresponding to Human ENPP2/ATX aa 300-400 conjugated to keyhole limpet haemocyanin.
Database link: Q13822
- WB: MOLT4, HeLa, HepG2 and Y79 whole cell lysates. IHC-P: FFPE human tonsil tissue sections. ICC/IF: U-87 MG cells.
This product was previously labelled as ENPP2
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
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...
PurityImmunogen affinity purified
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
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.
|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.|
FunctionHydrolyzes 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 specificityPredominantly 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 similaritiesBelongs to the nucleotide pyrophosphatase/phosphodiesterase family.
Contains 2 SMB (somatomedin-B) domains.
modificationsN-glycosylation, but not furin-cleavage, plays a critical role on secretion and on lysoPLD activity.
Cellular localizationSecreted. Secreted by most body fluids including serum and CSF. Also by adipocytes and numerous cancer cells.
- Information by UniProt
- ATX antibody
- ATX X antibody
- Autotaxin antibody
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.
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.
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.
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.