Anti-Osteopontin antibody (ab63856)
Key features and details
- Rabbit polyclonal to Osteopontin
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-Osteopontin antibody
See all Osteopontin primary antibodies -
Description
Rabbit polyclonal to Osteopontin -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
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Positive control
- This antibody gave a positive signal in the following lysates: Rat Skeletal Muscle Tissue Rat Brain Tissue HL60 Whole Cell Mouse Brain Tissue Rat Liver Tissue (data not shown) Mouse Liver Tissue (data not shown) Human Liver Tissue (data not shown)
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab63856 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 ICC/IF Use a concentration of 5 µg/ml. IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. WB Use a concentration of 1 µg/ml. Detects a band of approximately 32 kDa (predicted molecular weight: 32 kDa). Target
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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
modificationsExtensively phosphorylated on clustered serine residues.
N- and O-glycosylated.
Phosphorylation sites are present in the extracelllular medium. -
Cellular localization
Secreted. - Information by UniProt
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Database links
- Entrez Gene: 6696 Human
- Entrez Gene: 20750 Mouse
- Entrez Gene: 25353 Rat
- Omim: 166490 Human
- SwissProt: P10451 Human
- SwissProt: P10923 Mouse
- SwissProt: P08721 Rat
- Unigene: 313 Human
see all -
Alternative names
- BNSP antibody
- Bone sialoprotein 1 antibody
- Bone sialoprotein I antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody (ab63856)IHC image of Osteopontin staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab63856, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
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Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody (ab63856)ICC/IF image of ab63856 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 (ab63856, 5µg/ml) 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. -
Western blot - Anti-Osteopontin antibody (ab63856)All lanes : Anti-Osteopontin antibody (ab63856) at 1 µg/ml
Lane 1 : Skeletal Muscle (Rat) Tissue Lysate - normal tissue (ab29376)
Lane 2 : Brain (Rat) Tissue Lysate - normal tissue
Lane 3 : HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate (Human) Whole Cell Lysate
Lane 4 : Brain (Mouse) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 32 kDa
Additional bands at: 100 kDa, 42 kDa, 56 kDa, 88 kDa. We are unsure as to the identity of these extra bands.
Protocols
Datasheets and documents
References (10)
ab63856 has been referenced in 10 publications.
- Bhat OM et al. Medial calcification in the arterial wall of smooth muscle cell-specific Smpd1 transgenic mice: A ceramide-mediated vasculopathy. J Cell Mol Med 24:539-553 (2020). PubMed: 31743567
- Kouketsu A et al. Octacalcium phosphate collagen composite stimulates the expression and activity of osteogenic factors to promote bone regeneration. J Tissue Eng Regen Med 14:99-107 (2020). PubMed: 31721475
- Betriu N et al. Culture and Differentiation of Human Hair Follicle Dermal Papilla Cells in a Soft 3D Self-Assembling Peptide Scaffold. Biomolecules 10:N/A (2020). PubMed: 32354097
- Zhang L et al. Role of the Balance of Akt and MAPK Pathways in the Exercise-Regulated Phenotype Switching in Spontaneously Hypertensive Rats. Int J Mol Sci 20:N/A (2019). PubMed: 31766280
- Zhang Y et al. Guanine and nucleotide binding protein 3 promotes odonto/osteogenic differentiation of apical papilla stem cells via JNK and ERK signaling pathways. Int J Mol Med 43:382-392 (2019). PubMed: 30431055
- Song SZ et al. Targeting of SPP1 by microRNA-340 inhibits gastric cancer cell epithelial-mesenchymal transition through inhibition of the PI3K/AKT signaling pathway. J Cell Physiol N/A:N/A (2019). PubMed: 30953349
- Yan Y et al. Effect of Thyrotropin on Osteopontin, Integrin avß3, and VCAM-1 in the Endothelium via Activation of Akt. Int J Mol Sci 17:N/A (2016). PubMed: 27657042
- Koczorowska MM et al. Fibroblast activation protein-a, a stromal cell surface protease, shapes key features of cancer associated fibroblasts through proteome and degradome alterations. Mol Oncol 10:40-58 (2016). PubMed: 26304112
- Kim MO et al. Electromagnetic fields and nanomagnetic particles increase the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells. Int J Mol Med 35:153-60 (2015). PubMed: 25352086
- Nishimura I et al. Effect of osteogenic differentiation medium on proliferation and differentiation of human mesenchymal stem cells in three-dimensional culture with radial flow bioreactor. Regen Ther 2:24-31 (2015). PubMed: 31245456
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody (ab63856)IHC image of Osteopontin staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab63856, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
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Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody (ab63856)ICC/IF image of ab63856 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 (ab63856, 5µg/ml) 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.
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Western blot - Anti-Osteopontin antibody (ab63856)All lanes : Anti-Osteopontin antibody (ab63856) at 1 µg/ml
Lane 1 : Skeletal Muscle (Rat) Tissue Lysate - normal tissue (ab29376)
Lane 2 : Brain (Rat) Tissue Lysate - normal tissue
Lane 3 : HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate (Human) Whole Cell Lysate
Lane 4 : Brain (Mouse) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 32 kDa
Additional bands at: 100 kDa, 42 kDa, 56 kDa, 88 kDa. We are unsure as to the identity of these extra bands.
