Overview

  • Product name

    Anti-ENPP1/PC1 antibody [EPR22262-22]
    See all ENPP1/PC1 primary antibodies
  • Description

    Rabbit monoclonal [EPR22262-22] to ENPP1/PC1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human ENPP1/PC1 aa 1-100. The exact sequence is proprietary.
    Database link: P22413

  • Positive control

    • WB: Human kidney tissue lysate; MDA-MB-231, HepG2 and Huh7 whole cell lysate. IHC-P: Human pancreas and liver tissue. ICC/IF: HepG2 and MDA-MB-231 cells. Flow: HepG2 cells. IP: MDA-MB-231 whole cell lysate.
  • General notes

     This product was previously labelled as ENPP1

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab223268 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 1/1000. Detects a band of approximately 130 kDa (predicted molecular weight: 105 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

ICC/IF 1/100.
Flow Cyt 1/50.
IP 1/30.

Target

  • Function

    Involved primarily in ATP hydrolysis at the plasma membrane. Plays a role in regulating pyrophosphate levels, and functions in bone mineralization and soft tissue calcification. In vitro, has a broad specificity, hydrolyzing other nucleoside 5' triphosphates such as GTP, CTP, TTP and UTP to their corresponding monophosphates with release of pyrophosphate and diadenosine polyphosphates, and also 3',5'-cAMP to AMP. May also be involved in the regulation of the availability of nucleotide sugars in the endoplasmic reticulum and Golgi, and the regulation of purinergic signaling. Appears to modulate insulin sensitivity.
  • Tissue specificity

    Expressed in plasma cells and also in a number of non-lymphoid tissues, including the distal convoluted tubule of the kidney, chondrocytes and epididymis.
  • Involvement in disease

    Defects in ENPP1 are a cause of increased susceptibility for ossification of the posterior longitudinal ligament of the spine (OPLL) [MIM:602475]. OPLL is a common form of human myelopathy with a prevalence of as much as 4% in a variety of ethnic groups.
    Defects in ENPP1 are the cause of arterial calcification of infancy, generalized, type 1 (GACI1) [MIM:208000]. A severe autosomal recessive disorder characterized by calcification of the internal elastic lamina of muscular arteries and stenosis due to myointimal proliferation. The disorder is often fatal within the first 6 months of life because of myocardial ischemia resulting in refractory heart failure.
    Defects in ENPP1 are associated with obesity, glucose intolerance, and type II diabetes non-insulin dependent (NIDDM) [MIM:125853].
    Defects in ENPP1 are the cause of rickets hypophosphatemic autosomal recessive type 2 (ARHR2) [MIM:613312]. ARHR2 is a hereditary form of hypophosphatemic rickets, a disorder of proximal renal tubule function that causes phosphate loss, hypophosphatemia and skeletal deformities, including rickets and osteomalacia unresponsive to vitamin D. Symptoms are bone pain, fractures and growth abnormalities.
  • Sequence similarities

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

    The di-leucine motif is required for basolateral targeting in epithelial cells, and for targeting to matrix vesicles derived from mineralizing cells.
  • Post-translational
    modifications

    Autophosphorylated as part of the catalytic cycle of phosphodiesterase/pyrophosphatase activity.
    N-glycosylated.
    It has been suggested that the active SMB domain may be permitted considerable disulfide bond heterogeneity or variability, thus two alternate disulfide patterns based on 3D structures are described with 1 disulfide bond conserved in both.
  • Cellular localization

    Membrane. Basolateral cell membrane. Targeted to the basolateral membrane in polarized epithelial cells and in hepatocytes, and to matrix vesicles in osteoblasts. In bile duct cells and cancer cells, located to the apical cytoplasmic side.
  • Information by UniProt
  • Database links

  • Alternative names

    • Alkaline phosphodiesterase 1 antibody
    • ARHR2 antibody
    • COLED antibody
    • E-NPP 1 antibody
    • Ectonucleotide pyrophosphatase/phosphodiesterase 1 antibody
    • Ectonucleotide pyrophosphatase/phosphodiesterase family member 1 antibody
    • ENPP1 antibody
    • ENPP1_HUMAN antibody
    • Ly 41 antigen antibody
    • M6S1 antibody
    • Membrane component chromosome 6 surface marker 1 antibody
    • NPP1 antibody
    • NPPase antibody
    • NPPS antibody
    • Nucleotide pyrophosphatase antibody
    • PC 1 antibody
    • PC-1 antibody
    • PCA1 antibody
    • PDNP1 antibody
    • Phosphodiesterase I/nucleotide pyrophosphatase 1 antibody
    • Plasma cell membrane glycoprotein 1 antibody
    • Plasma-cell membrane glycoprotein PC-1 antibody
    see all

Images

  • All lanes : Anti-ENPP1/PC1 antibody [EPR22262-22] (ab223268) at 1/1000 dilution

    Lane 1 : Human kidney tissue lysate
    Lane 2 : MDA-MB-231 (human breast adenocarcinoma cell line) whole cell lysate
    Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 4 : Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
    Lanes 2-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 105 kDa
    Observed band size: 130 kDa
    why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST 

    Exposure time:

    Lane 1:3 minutes;
    Lane 2:70 seconds;
    Lane 3:15 seconds;
    Lane 4:3 minutes

    The molecular weight observed is consistent with what has been described in the literature (PMID: 19577557).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cells labeling ENPP1/PC1 (green) with ab223268 at 1/100 dilution, followed by Alexa Fluor® 488 Goat anti-Rabbit secondary antibody (ab150077) at 1/1000 dilution. Confocal image showing membranous staining in HepG2 cells. Tubulin was detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red). The nuclear countertsain was DAPI (Blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Alexa Fluor® 488 Goat anti-Rabbit (ab150077) at 1/200 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MDA-MB-231 (human breast adenocarcinoma cell line) cells labeling ENPP1/PC1 (green) with ab223268 at 1/100 dilution, followed by Alexa Fluor® 488 Goat anti-Rabbit secondary antibody (ab150077) at 1/1000 dilution. Confocal image showing membranous staining in MDA-MB-231 cells. Tubulin was detected using Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (Red). The nuclear countertsain was DAPI (Blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Alexa Fluor® 488 Goat anti-Rabbit (ab150077) at 1/200 dilution.

  • ENPP1/PC1 was immunoprecipitated from 0.35 mg of MDA-MB-231 (human breast adenocarcinoma cell line) whole cell lysate using ab223268 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab223268 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

    Lane 1: MDA-MB-231 whole cell lysate 10μg (input)
    Lane 2: ab223268 IP in MDA-MB-231 whole cell lysate (+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab223268 in MDA-MB-231 whole cell lysate (-).

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 30 seconds.

     

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cells labeling ENPP1/PC1 with ab223268 (red) compared with a Rabbit monoclonal IgG  isotype control (ab172730) (black)and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue stained for ENPP1/PC1 using ab223268 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining on human liver (PMID: 25539584; PMID: 9344668; PMID: 23861746) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue stained for ENPP1/PC1 using ab223268 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining on human pancreas (PMID: 9344668; PMID: 23861746) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

References

ab223268 has not yet been referenced specifically in any publications.

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