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

  • 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
    • 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

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
    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
  • 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

This product has been referenced in:
  • Bai G  et al. Synthesis of a Cleaved Form of Osteopontin by THP-1 Cells and Its Alteration by Phorbol 12-Myristate 13-Acetate and BCG Infection. Int J Mol Sci 19:N/A (2018). Read more (PubMed: 29385060) »
  • Choi Y  et al. Preventive effects of indole-3-carbinol against alcohol-induced liver injury in mice via antioxidant, anti-inflammatory, and anti-apoptotic mechanisms: Role of gut-liver-adipose tissue axis. J Nutr Biochem 55:12-25 (2018). Mouse . Read more (PubMed: 29331880) »

See all 90 Publications for this product

Customer reviews and Q&As

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Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Nov 28 2017

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

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Kidney)
Loading amount
150 µg
Specification
Kidney
Treatment
12w after kidney transplantation
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C
Username

Mr. Ting Zhang

Verified customer

Submitted Sep 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Developing skeleton)
Specification
Developing skeleton
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 18 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Kidney)
Specification
Kidney
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Username

Mrs. Judith Kranz

Verified customer

Submitted Feb 18 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (jaw bone (mandible) with teeth)
Specification
jaw bone (mandible) with teeth
Fixative
Paraformaldehyde
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: 0.4% pepsin 37°C
Permeabilization
No
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 4% · Temperature: rt°C
Username

Prof. Werner Goetz

Verified customer

Submitted Sep 05 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Breast)
Specification
Breast
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 10%
Username

Abcam user community

Verified customer

Submitted Feb 16 2007

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