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    osteopontin-antibody-epr21139-316-ab214050.pdf

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Cardiovascular Angiogenesis Adhesion / ECM Extracellular Matrix
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RecombinantRabMAb

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

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)
  • Immunoprecipitation - Anti-Osteopontin antibody [EPR21139-316] (ab214050)
  • Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)
  • 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)
  • Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody [EPR21139-316] (ab214050)
  • Anti-Osteopontin antibody [EPR21139-316] (ab214050)

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, ICC/IF
  • Reacts with: Human, Recombinant fragment

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free

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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, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Human, Recombinant fragment
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR21139-316
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Angiogenesis
    • Adhesion / ECM
    • Extracellular Matrix
    • Signal Transduction
    • Cytoskeleton / ECM
    • Extracellular Matrix
    • Structures
    • Bone
    • Stem Cells
    • Mesenchymal Stem Cells
    • Osteogenesis
    • Cancer
    • Tumor biomarkers
    • Other

Associated products

  • Alternative Versions

    • Anti-Osteopontin antibody (ab11503)
    • Anti-Osteopontin antibody [EPR21139-316] - BSA and Azide free (ab236213)
    • Alexa Fluor® 594 Anti-Osteopontin antibody [EPR21139-316] (ab282279)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human Osteopontin protein (ab92964)
  • Related Products

    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

The Abpromise guarantee

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).
ICC/IF
1/50.
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).
ICC/IF
1/50.

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.
  • Target information above from: UniProt accession P10451 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6696 Human
    • Omim: 166490 Human
    • SwissProt: P10451 Human
    • Unigene: 313 Human
    • 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody [EPR21139-316] (ab214050)
      Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody [EPR21139-316] (ab214050)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-87 MG cell line labelling Osteopontin with ab214050 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplamic staining in U-87 MG cell line. The nuclear counter stain is DAPI (blue).

      The negative controls is as follows:
      -ve control: Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

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

    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

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (9)

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

    ab214050 has been referenced in 9 publications.

    • Zhao L  et al. Catalpol Protects Against High Glucose-Induced Bone Loss by Regulating Osteoblast Function. Front Pharmacol 12:626621 (2021). PubMed: 33776769
    • Yang L  et al. Wnt7a promotes the osteogenic differentiation of human mesenchymal stem cells. Int J Mol Med 47:N/A (2021). PubMed: 33846764
    • Li M  et al. Calculus-related functional protein expression in ureteral calculus-adhered polyp: A preliminary study. Medicine (Baltimore) 100:e26512 (2021). PubMed: 34160472
    • Gao X  et al. LINC00899 promotes osteogenic differentiation by targeting miR-374a and RUNX2 expression. Exp Ther Med 22:1071 (2021). PubMed: 34447464
    • Xiang J  et al. lncRNA SNHG1 attenuates osteogenic differentiation via the miR-101/DKK1 axis in bone marrow mesenchymal stem cells. Mol Med Rep 22:3715-3722 (2020). PubMed: 32901867
    • Liu Z  et al. Proteomic Analysis of Human Endometrial Tissues Reveals the Roles of PI3K/AKT/mTOR Pathway and Tumor Angiogenesis Molecules in the Pathogenesis of Endometrial Cancer. Biomed Res Int 2020:5273969 (2020). PubMed: 32908897
    • Wangzhou K  et al. MiR-143-3p Inhibits Osteogenic Differentiation of Human Periodontal Ligament Cells by Targeting KLF5 and Inactivating the Wnt/ß-Catenin Pathway. Front Physiol 11:606967 (2020). PubMed: 33603676
    • Zhu L  et al. Hyperhomocysteinemia induces vascular calcification by activating the transcription factor RUNX2 via Krüppel-like factor 4 up-regulation in mice. J Biol Chem 294:19465-19474 (2019). PubMed: 31628194
    • 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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