Recombinant
RabMAb

Recombinant Anti-Osteopontin antibody [EPR21139-316] (ab214050)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21139-316] to Osteopontin
  • Suitable for: IP, IHC-P, WB
  • Reacts with: Human

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

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

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

    • Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)
      Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)
      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).

    • Immunoprecipitation - Anti-Osteopontin antibody [EPR21139-316] (ab214050)
      Immunoprecipitation - Anti-Osteopontin antibody [EPR21139-316] (ab214050)

      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).

    • Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)
      Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21139-316] (ab214050)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21139-316] (ab214050)

      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).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21139-316] (ab214050)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21139-316] (ab214050)

      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.

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References

    Publishing research using ab214050? Please let us know so that we can cite the reference in this datasheet.

    ab214050 has been referenced in 1 publication.

    • Chen P  et al. Identification and validation of four hub genes involved in the plaque deterioration of atherosclerosis. Aging (Albany NY) 11:6469-6489 (2019). PubMed: 31449494

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