Anti-ENPP2/ATX antibody [1F8] (ab77104)
Key features and details
- Mouse monoclonal [1F8] to ENPP2/ATX
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-ENPP2/ATX antibody [1F8]
See all ENPP2/ATX primary antibodies -
Description
Mouse monoclonal [1F8] to ENPP2/ATX -
Host species
Mouse -
Tested applications
Suitable for: IHC-P, WBmore details
Unsuitable for: ICC -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse -
Immunogen
Recombinant full length protein corresponding to Human ENPP2/ATX.
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Positive control
- WB: HEK293 whole cell lysate and in the following human tissue lysates: kidney; placenta; ovary; small intestine. IHC: human tonsil paraffin sections.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
1F8 -
Isotype
IgG1 -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab77104 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 |
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IHC-P |
Use a concentration of 1 µg/ml.
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WB |
Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 99 kDa).
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Notes |
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IHC-P
Use a concentration of 1 µg/ml. |
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 99 kDa). |
Target
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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
modificationsN-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
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Database links
- Entrez Gene: 5168 Human
- Entrez Gene: 18606 Mouse
- Omim: 601060 Human
- SwissProt: Q13822 Human
- SwissProt: Q9R1E6 Mouse
- Unigene: 190977 Human
- Unigene: 250256 Mouse
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Alternative names
- ATX antibody
- ATX X antibody
- Autotaxin antibody
see all
Images
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All lanes : Anti-ENPP2/ATX antibody [1F8] (ab77104) at 1/1000 dilution
Lane 1 : HEK-293 cell lysate at 20 µg
Lane 2 : PANC-1 cell lysate at 20 µg
Lane 3 : U-87 MG cell lysate at 20 µg
Lane 4 : Huh7 cell lysate at 20 µg
Lane 5 : HepG2 cell lysate at 20 µg
Lane 6 : Wild Type HeLa cell lysate at 20 µg
Lane 7 : ENPP2 knockout HeLa cell lysate at 20 µg
Lane 8 : Empty cell lysate at 0 µg
Lane 9 : Recombinant Human ENPP-2/Autotaxin Protein, CF cell lysate at 0.5 µg
Lane 10 : Recombinant Human ENPP2/ATX Protein ab126918 cell lysate at 0.5 µg
Performed under reducing conditions.
Predicted band size: 99 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?Anti-ENPP2 antibody [1F8] (ab77104) staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab77104 was shown to bind specifically to ENPP2. A band was observed at 105 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in ENPP2 knockout cell line. To generate this image, wild-type and ENPP2 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
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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
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All lanes : Anti-ENPP2/ATX antibody [1F8] (ab77104) at 10 µg/ml
Lane 1 : Human kidney tissue lysate - total protein (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 -
All lanes : Anti-ENPP2/ATX antibody [1F8] (ab77104) at 10 µg/ml
Lane 1 : Human placenta tissue lysate - total protein (ab29745)
Lane 2 : Human ovary tissue lysate - total protein (ab30222)
Lane 3 : Human small intestine tissue lysate - total protein (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.
Performed under reducing conditions.
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 minutesENPP2/ATX contains three potential glycosylation sites (SwissProt), which might explain its migration at a higher molecular weight than predicted.
Datasheets and documents
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Datasheet download
References (10)
ab77104 has been referenced in 10 publications.
- Trovato FM et al. Lysophosphatidylcholines modulate immunoregulatory checkpoints in peripheral monocytes and are associated with mortality in people with acute liver failure. J Hepatol 78:558-573 (2023). PubMed: 36370949
- Cao H et al. Autotaxin signaling facilitates β cell dedifferentiation and dysfunction induced by Sirtuin 3 deficiency. Mol Metab 60:101493 (2022). PubMed: 35398277
- Bhattarai S et al. The ATX-LPA Axis Regulates Vascular Permeability during Cerebral Ischemic-Reperfusion. Int J Mol Sci 23:N/A (2022). PubMed: 35456953
- Amaral RF et al. Microglial lysophosphatidic acid promotes glioblastoma proliferation and migration via LPA1 receptor. J Neurochem 156:499-512 (2021). PubMed: 32438456
- Deng W et al. Hepatitis B Virus Promotes Hepatocellular Carcinoma Progression Synergistically With Hepatic Stellate Cells via Facilitating the Expression and Secretion of ENPP2. Front Mol Biosci 8:745990 (2021). PubMed: 34805271
- Liu S et al. Lysophosphatidic acid mediated PI3K/Akt activation contributed to esophageal squamous cell cancer progression. Carcinogenesis 42:611-620 (2021). PubMed: 33367557
- Kaya B et al. Lysophosphatidic Acid-Mediated GPR35 Signaling in CX3CR1+ Macrophages Regulates Intestinal Homeostasis. Cell Rep 32:107979 (2020). PubMed: 32755573
- Xie D et al. The FOXM1/ATX signaling contributes to pancreatic cancer development. Am J Transl Res 12:4478-4487 (2020). PubMed: 32913521
- Zhang G et al. ATX-LPA axis facilitates estrogen-induced endometrial cancer cell proliferation via MAPK/ERK signaling pathway. Mol Med Rep 17:4245-4252 (2018). PubMed: 29328374
- Su SC et al. Autotaxin-lysophosphatidic acid signaling axis mediates tumorigenesis and development of acquired resistance to sunitinib in renal cell carcinoma. Clin Cancer Res 19:6461-72 (2013). IHC-P ; Human . PubMed: 24122794