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    osteopontin-antibody-ab8448.pdf

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Cardiovascular Angiogenesis Adhesion / ECM Extracellular Matrix
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Anti-Osteopontin antibody (ab8448)

  • Datasheet
  • SDS
Reviews (12)Q&A (18)References (235)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody (ab8448)
  • Western blot - Anti-Osteopontin antibody (ab8448)
  • Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody (ab8448)
  • Cellular activity - Anti-Osteopontin antibody (ab8448)

Key features and details

  • Rabbit polyclonal to Osteopontin
  • Suitable for: IHC-P, ICC/IF, WB
  • Reacts with: Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Osteopontin antibody [EPR21139-316] (ab214050)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-Osteopontin antibody
    See all Osteopontin primary antibodies
  • Description

    Rabbit polyclonal to Osteopontin
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human Osteopontin aa 170-183 conjugated to keyhole limpet haemocyanin.
    Sequence:

    CKSKKFRRPDIQYPD


    Database link: P10451
    (Peptide available as ab9078)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

Properties

  • 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

    Preservative: 0.1% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
  • Concentration information loading...
  • Purity

    Whole antiserum
  • Clonality

    Polyclonal
  • 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

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunohistochemistry kits

    • BrdU Immunohistochemistry Kit (ab125306)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Osteopontin protein (ab92964)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab8448 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IHC-P
Human
WB
Human
Application Abreviews Notes
IHC-P (5)
1/100 - 1/300.
ICC/IF
1/1000.
WB (5)
1/1000. at this dilution, the antibody will strongly detect approximately 250 ng of OPN protein on a blot.
Notes
IHC-P
1/100 - 1/300.
ICC/IF
1/1000.
WB
1/1000. at this dilution, the antibody will strongly detect approximately 250 ng of OPN protein on a blot.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody (ab8448)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody (ab8448)

      Human Breast tumour section. Osteopontin is a normal component of elastic fibers in the breast (shown heavily stained in this section). There is also weak staining of the extracellular matrix. Osteopontin is not believed to be expressed inside breast tumour cells, and there is no staining in the intracellular region of the breast cells in this section.

      Osteopontin antibody (ab8448) used at 1:100-1:300. No antigen retrieval is required.

    • Western blot - Anti-Osteopontin antibody (ab8448)
      Western blot - Anti-Osteopontin antibody (ab8448)
      All lanes : Anti-Osteopontin antibody (ab8448) at 1/1000 dilution

      Lane 2 : Human Osteopontin
      Lane 3 : MMP-cleaved Human Osteopontin

      Lysates/proteins at 0.25 µg per lane.

      Secondary
      All lanes : HRP-conjugated Goat anti-Rabbit IgG at 1/10000 dilution


      The osteopontin antibody (ab8448) is used at 1:1000 dilution on a blot with 250ng human osteopontin (lane 2)and MMP-cleaved osteopontin (lane 3
    • Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody (ab8448)
      Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody (ab8448)
      ICC/IF image of ab8448 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8448, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Cellular activity - Anti-Osteopontin antibody (ab8448)
      Cellular activity - Anti-Osteopontin antibody (ab8448)This image is courtesy of Agnihotri et al, JBC 2001
      OPN is cleaved by MMP to yield 2 fragments, which migrate at 40kD(N terminal) and 32kD (C terminal). The C terminal fragment can undergo further cleavage by both of these MMPs (see Agnihotri et al, JBC 2001 for further details). The epitope recognised by ab8448 is shown in violet. This antibody detects the full length OPN and the 32kD fragment. It does not recognise the 40kD fragment.

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (235)

    Publishing research using ab8448? Please let us know so that we can cite the reference in this datasheet.

    ab8448 has been referenced in 235 publications.

    • Li L  et al. XIST/miR-376c-5p/OPN axis modulates the influence of proinflammatory M1 macrophages on osteoarthritis chondrocyte apoptosis. J Cell Physiol 235:281-293 (2020). PubMed: 31215024
    • Geng Y  et al. Overexpression of mechanical sensitive miR-337-3p alleviates ectopic ossification in rat tendinopathy model via targeting IRS1 and Nox4 of tendon-derived stem cells. J Mol Cell Biol 12:305-317 (2020). PubMed: 31065679
    • Li P  et al. Loss of PARP-1 attenuates diabetic arteriosclerotic calcification via Stat1/Runx2 axis. Cell Death Dis 11:22 (2020). PubMed: 31924749
    • Marycz K  et al. Iron oxides nanoparticles (IOs) exposed to magnetic field promote expression of osteogenic markers in osteoblasts through integrin alpha-3 (INTa-3) activation, inhibits osteoclasts activity and exerts anti-inflammatory action. J Nanobiotechnology 18:33 (2020). PubMed: 32070362
    • Gadalla D & Goldstein AS Improving the Osteogenicity of PCL Fiber Substrates by Surface-Immobilization of Bone Morphogenic Protein-2. Ann Biomed Eng 48:1006-1015 (2020). PubMed: 31115719
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 30 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Osteopontin antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Bone marrow)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6
    Specification
    Bone marrow
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More

    Dr. Queralt Vallmajó

    Verified customer

    Submitted Mar 18 2020

    Western blot abreview for Anti-Osteopontin antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Pig Tissue lysate - whole (heart)
    Gel Running Conditions
    Reduced Denaturing (4-12% bis tris)
    Loading amount
    20 µg
    Specification
    heart
    Blocking step
    LIcor block buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 37°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 28 2017

    Western blot abreview for Anti-Osteopontin antibody

    Poor
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Established cell line)
    Gel Running Conditions
    Reduced Denaturing (Tris Glycine, Gradient gels of 4-20%)
    Loading amount
    30 µg
    Specification
    Established cell line
    Blocking step
    Milk as blocking agent for 8 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
    Read More

    Anantha Marisetty

    Verified customer

    Submitted Mar 10 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Osteopontin antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 35°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate buffer
    Sample
    Mouse Tissue sections (Femur)
    Specification
    Femur
    Permeabilization
    No
    Fixative
    Paraformaldehyde
    Read More

    Mr. Helder Fonseca

    Verified customer

    Submitted Oct 06 2014

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Osteopontin antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Antigen retrieval step
    None
    Sample
    Human Tissue sections (Tooth)
    Specification
    Tooth
    Permeabilization
    No
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 28 2014

    Western blot abreview for Anti-Osteopontin antibody

    Below Average
    Abreviews
    Abreviews
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Rat Tissue lysate - whole (Liver, WAT)
    Specification
    Liver, WAT
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Read More

    Dr. Elisa Fernandez

    Verified customer

    Submitted Feb 26 2014

    Question

    Inquiry: Hi, I'm wondering if this antibody (ab11503) binds only the full length molecule or if it is able to bind even the cleaved fragments (as does ab8448). Thank you

    Read More

    Abcam community

    Verified customer

    Asked on Jul 09 2013

    Answer

    The immunogen of antibody is from amino acid 170-183 of human Osteopontin http://www.uniprot.org/uniprot/P10451. The antibody should bind to full length protein as well the immunogen sequence containing fragment.

    Read More

    Padamjeet Singh

    Abcam Scientific Support

    Answered on Jul 09 2013

    Question

    Inhibits this antibody the osteopontin? Works well in ICC.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 09 2013

    Answer

    Vielen Dank für Ihren Anruf.

    Leider wurde es nochnoch nicht untersucht, ob dieser Antikörper das Osteopontin hemmen kann.

    Ich vermute allerdings, dass dies nicht der Fall sein wird, da das Immunogen nicht gegen das Motiv "Cell attachment site" in Position AS 159-161 gerichtet ist. Dies bedeutet, dass das Protein wahrscheinlich weiterhin die Zellen binden kann, um eine Wirkung zu entfalten.

    Falls Sie dies allerdings dennoch ausprobieren möchten (da Sie den Antikörper ja bereits verwenden), rate ich ihnen auf jeden Fall vorher das Natriumazide aus dem Antikörper zu entfernen, da dieses toxisch auf Ihre Zellen wirkt.

    Ich hoffe, dies hilft Ihnen trotzdem weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

    Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Jan 09 2013

    Question

    Inquiry: I was wondering if the raabit polyclonal OPN antibody ab14176 has been used in western blotting before, if so, which OPN bands would it detect? I know a previous one we had ab8448 detects both full size OPN at 66 kDa and MMP cleaved at 32 kDa. I was wondering if ab14176 is the same as we are seeing bands at similar regions. Thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Dec 17 2012

    Answer

    Thank you very much for your interest in our products.

    I can confirm that we have not tested ab14176 in WB and therefore do not have this information determined by an experiment. But since the immunogen of ab14176 is in the same reagion as the immunogen of ab8448 we would also expect similar WB results.

    We do not currently have an image of ab14176 in WB.

    Simply follow these easy steps below to apply for our AbTrial Program:

    1. Reply to this email, letting us know you are interested in submitting an abreview with an image in WB for ab14176.

    2. Our scientists will email you an inactive personal discount code for the value of the product.

    3. Prepare the abreview and image.

    4. Submit an image of your results, including your discount code in the additional notes section of your Abreview.

    5. Once the Abreview is submitted, the discount code will become active.

    6. Apply your discount code on your next order to receive that value off.

    Please let me know if you have any questions about this offer and I would be happy to help you further.

    The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

    Read More

    Abcam Scientific Support

    Answered on Dec 17 2012

    Question

    I am answering the question on the technical issue:

    1. Can you please tell me the species and tissue type or cell type of the sample and how it was prepared? For example, was it prepared with protease inhibitors and was is denatured and reduced before loading into the gel?

    mouse, adipose tissue, RIPA buffer lysate with protease inhibitor, denatured with beta Mercaptoethanol, 50ug loading,


    2. How was the membrane blocked (milk, BSA, etc.) and how much antibody did you incubate the membrane in?

    Blocked with 5% milk, overnight incubation at 4 degree,


    3. Finally, could you please describe what the stain of the blot looked like? Where did you see bands, if any?

    multiple bands

    Read More

    Abcam community

    Verified customer

    Asked on Nov 15 2012

    Answer

    Thank you for sending these protocol details. The antibody is expected to detect multiple bands representing processed osteopontin, as described in the note and the diagram on the datasheet,

    OPN is cleaved by MMP to yield 2 fragments, which migrate at 40kD(N terminal) and 32kD (C terminal). The C terminal fragment can undergo further cleavage by both of these MMPs (see Agnihotri et al, JBC 2001 for further details). The epitope recognised by ab8448 is shown in violet. This antibody detects the full length OPN and the 32kD fragment. It does not recognise the 40kD fragment.

    Click here (or use the following: https://www.abcam.com/Osteopontin-antibody-ab8448.html).

    Do the bands you see correspond to these sizes?

    Read More

    Abcam Scientific Support

    Answered on Nov 15 2012

    1-10 of 30 Abreviews or Q&A

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