Anti-Osteopontin antibody (ab8448)

Reviews (11)Q&A (18)References (120)

Overview

  • Product name
    Anti-Osteopontin antibody
    See all Osteopontin primary antibodies
  • Description
    Rabbit polyclonal to Osteopontin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, ICC/IF, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
  • 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

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
    see all
  • 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody (ab8448)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody (ab8448)Joshi, S. et al PLoS One. 2017 Nov 1;12(11):e0185009. doi: 10.1371/journal.pone.0185009. eCollection 2017.

    Immuno-histological staining of EG treated rat kidney

    Formalin-fixed, paraffin-embedded rat kidney tissue stained for Osteopontin using ab8448 at 1/100 dilution in immunohistochemical analysis.

    A) Control B) on Day 14 showing light staining of the tubular epithelial cells, shown by black arrows and C) on Day 28 showing strong staining of renal tubular epithelial cells as well as tubular contents (Black arrows), Magnification, X 40, Scale bar 100μm.

  • 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.

  • Immunohistochemistry (Frozen sections) - Anti-Osteopontin antibody (ab8448)
    Immunohistochemistry (Frozen sections) - Anti-Osteopontin antibody (ab8448)This image is a courtesy of Anonymous Abreview

    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

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

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody (ab8448)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody (ab8448)Image is courtesy of an Abreview submitted by Helder Fonseca

    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

  • Other - Anti-Osteopontin antibody (ab8448)
    Other - 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.

References

This product has been referenced in:
  • Oryan A  et al. Synergistic effect of strontium, bioactive glass and nano-hydroxyapatite promotes bone regeneration of critical-sized radial bone defects. J Biomed Mater Res B Appl Biomater 107:50-64 (2019). Read more (PubMed: 29468802) »
  • Yan Y  et al. Vascularized 3D printed scaffolds for promoting bone regeneration. Biomaterials 190-191:97-110 (2019). Read more (PubMed: 30415019) »
See all 120 Publications for this product

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

Customer reviews and Q&As

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1-10 of 29 Abreviews or Q&A

Western blot abreview for Anti-Osteopontin antibody

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

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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.

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Question

Inhibits this antibody the osteopontin? Works well in ICC.

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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

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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,

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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.

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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

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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/index.html?datasheet=8448).

Do the bands you see correspond to these sizes?

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Question

Inquiry: Sehr geehrte Damen und Herren, ich interessiere mich für die folgenden Produkte:
Anti-Osteopontin antibody (ab8448)
Anti-TRAF6 antibody (ab94769)
Anti-TGF beta 1 antibody [TB21] (ab27969)
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (FITC) (ab6717)
Könnten Sie mir bitte ein konkurrenzfähiges Angebot für diese Antikörper schicken? Mit freundlichen Grüßen

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Answer

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.

Sie koennen ganz einfach per E-mail bestellen: mailto:orders@abcam.com


Geben sie bei der Bestellung bitte den Aktionscode an: 4-FOR-3-EB3AB


Ich hoffe, dies hilft Ihnen 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

1-10 of 29 Abreviews or Q&A

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