Overview

  • Product name

    Anti-Osteopontin antibody [EPR21139-316]
    See all Osteopontin primary antibodies
  • Description

    Rabbit monoclonal [EPR21139-316] to Osteopontin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human Osteopontin aa 1-300. The exact sequence is proprietary.
    Database link: P10451

  • Positive control

    • WB: Human Osteopontin recombinant protein (aa17-300) produced in a mammalian expression system; Human urine and fetal kidney lysates. IHC-P: Human kidney and liver tissues. IP: Human kidney lysate.
  • General notes

    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 ab214050 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
IP 1/30.
IHC-P 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 45, 60, 70, 90 kDa (predicted molecular weight: 35 kDa).

Target

  • Function

    Binds tightly to hydroxyapatite. Appears to form an integral part of the mineralized matrix. Probably important to cell-matrix interaction.
    Acts as a cytokine involved in enhancing production of interferon-gamma and interleukin-12 and reducing production of interleukin-10 and is essential in the pathway that leads to type I immunity.
  • Tissue specificity

    Bone. Found in plasma.
  • Sequence similarities

    Belongs to the osteopontin family.
  • Post-translational
    modifications

    Extensively phosphorylated on clustered serine residues.
    N- and O-glycosylated.
    Phosphorylation sites are present in the extracelllular medium.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Database links

  • Alternative names

    • BNSP antibody
    • Bone sialoprotein 1 antibody
    • Bone sialoprotein I antibody
    • BSP I antibody
    • BSPI antibody
    • Early T lymphocyte activation 1 antibody
    • ETA 1 antibody
    • ETA1 antibody
    • MGC110940 antibody
    • Nephropontin antibody
    • OPN antibody
    • Osteopontin antibody
    • osteopontin/immunoglobulin alpha 1 heavy chain constant region fusion protein antibody
    • OSTP_HUMAN antibody
    • PSEC0156 antibody
    • secreted phosphoprotein 1 (osteopontin bone sialoprotein I early T lymphocyte activation 1) antibody
    • secreted phosphoprotein 1 (osteopontin, bone sialoprotein I, early T-lymphocyte activation 1) antibody
    • Secreted phosphoprotein 1 antibody
    • SPP 1 antibody
    • SPP-1 antibody
    • SPP1 antibody
    • SPP1/CALPHA1 fusion antibody
    • Urinary stone protein antibody
    • Uropontin antibody
    see all

Images

  • All lanes : Anti-Osteopontin antibody [EPR21139-316] (ab214050) at 1/1000 dilution

    Lane 1 : Human urine at 20 µg
    Lane 2 : Human fetal kidney lysate at 10 µg

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

    Predicted band size: 35 kDa
    Observed band size: 45,60,70,90 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    The multiple bands observed are likely due to expression of different isoforms, as well as glycosylation (PMID: 8761944; PMID: 11375993; PMID: 8535240; PMID: 12890718).

  • Osteopontin was immunoprecipitated from 0.35 mg of human kidney lysate with ab214050 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214050 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution

    Lane 1: Human kidney lysate 10 µg (Input). 

    Lane 2: ab214050 IP in human kidney lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214050 in human kidney lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    The multiple bands observed are likely due to expression of different isoforms, as well as glycosylation (PMID: 8761944; PMID: 11375993; PMID: 8535240; PMID: 12890718).

  • Anti-Osteopontin antibody [EPR21139-316] (ab214050) at 1/1000 dilution + Human Osteopontin recombinant protein (aa17-300) produced in a mammalian expression at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 35 kDa
    Observed band size: 60-65 kDa why is the actual band size different from the predicted?


    Exposure time: 26 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The his-tagged recombinant protein was produced in a mammalian expression system, thereby allowing it to be post-translationally modified.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Osteopontin with ab214050 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in bile ducts of human liver (PMID: 16221502). 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.

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Osteopontin with ab214050 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use. Cytoplasmic staining in distal tubules of human kidney (PMID: 17938278), performed on a Leica Biosystems BOND® RX instrument. 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.

    Perform heat-mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 minutes.

References

ab214050 has not yet been referenced specifically in any publications.

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