Overview

Properties

Applications

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.

Application Abreviews 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.
IHC-Fr 1/100 - 1/500. PubMed: 16128620

Target

  • FunctionBinds 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 specificityBone. Found in plasma.
  • Sequence similaritiesBelongs 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 localizationSecreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • BNSP antibody
    • Bone sialoprotein 1 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 antibody
    • SPP 1 antibody
    • SPP-1 antibody
    • SPP1 antibody
    • SPP1/CALPHA1 fusion antibody
    • Urinary stone protein antibody
    • Uropontin antibody
    see all

Anti-Osteopontin antibody images

  • 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
    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
  • 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.

  • ab8448 staining Osteopontin in mouse developing skeleton tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with paraformaldehyde before permeabilization with 0.1% Triton and blocking with 20% serum was performed for 1 hour at RT. The sample was incubated with primary antibody (1/200) in 20%FBS/PBS for 16 hours at 40C. An Alexa Fluor®488-conjugated donkey polyclonal to rabbit IgG was used as secondary antibody at 1/200 dilution. In the image: Red Rhodamine Phalloidin (muscle), Blue DAPI (nuclei), Green Osteopontin.

    See Abreview

  • 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.

  • ab8448 staining Osteopontin in Mouse femur tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% BSA for 1 hour at 35°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/50) for 14 hours at 4°C. An Alkaline Phosphatase-conjugated Goat anti-rabbit IgG F(ab')2 polyclonal (1/100) was used as the secondary antibody.

    See Abreview

  • 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.

References for Anti-Osteopontin antibody (ab8448)

This product has been referenced in:
  • Dang PN  et al. Controlled Dual Growth Factor Delivery From Microparticles Incorporated Within Human Bone Marrow-Derived Mesenchymal Stem Cell Aggregates for Enhanced Bone Tissue Engineering via Endochondral Ossification. Stem Cells Transl Med 5:206-17 (2016). Human . Read more (PubMed: 26702127) »
  • Spina A  et al. NZ-GMP Approved Serum Improve hDPSC Osteogenic Commitment and Increase Angiogenic Factor Expression. Front Physiol 7:354 (2016). ICC/IF ; Human . Read more (PubMed: 27594842) »

See all 40 Publications for this product

Product Wall

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
Username

Anantha Marisetty

Verified customer

Submitted Mar 10 2016

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
Username

Mr. Helder Fonseca

Verified customer

Submitted Oct 06 2014

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
Username

Abcam user community

Verified customer

Submitted Feb 28 2014

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
Username

Dr. Elisa Fernandez

Verified customer

Submitted Feb 26 2014

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.

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 ...

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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 t...

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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 ...

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Vielen Dank für Ihre Anfrage.

Ich freue mich ihnen mitteilen zu koennen dass wir fuer den gesamten November unsere Aktion 4 fuer 3 laufen haben. Bei der Bestellung von 4 Produkten bekommen Sie das guenstigste Produkt kostenfrei.

...

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Vielen Dank für Ihre Anfrage.

Leider ist es nicht möglich die Konzentration von ab8448 zu bestimmen.

Bei ab8448 handelt es sich um unaufgereinigtes Vollserum.

Die Konzentration von Antikörpern wird über...

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1-10 of 28 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"