Recombinant Anti-Osteopontin antibody [EPR21139-316] (ab214050)
![Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)](https://www.abcam.com/ps/products/214/ab214050/Images/ab214050-311746-anti-osteopontin-antibody-epr21139-316-western-blot.jpg "Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21139-316] to Osteopontin
- Suitable for: IP, IHC-P, WB, ICC/IF
- Reacts with: Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-Osteopontin antibody [EPR21139-316]
See all Osteopontin primary antibodies -
Description
Rabbit monoclonal [EPR21139-316] to Osteopontin -
Host species
Rabbit -
Tested applications
Suitable for: IP, IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human, Recombinant fragment -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human Osteopontin recombinant protein (aa17-300) produced in a mammalian expression system; Human urine and fetal kidney lysates. IHC-P: Human kidney and liver tissues. IP: Human kidney lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21139-316 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab214050 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 |
|---|---|---|
| IP |
1/30.
|
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| IHC-P |
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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| WB |
1/1000. Detects a band of approximately 45, 60, 70, 90 kDa (predicted molecular weight: 35 kDa).
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| ICC/IF |
1/50.
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| Notes |
|---|
|
IP
1/30. |
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IHC-P
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
|
WB
1/1000. Detects a band of approximately 45, 60, 70, 90 kDa (predicted molecular weight: 35 kDa). |
|
ICC/IF
1/50. |
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
- Omim: 166490 Human
- SwissProt: P10451 Human
- Unigene: 313 Human
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Alternative names
- BNSP antibody
- Bone sialoprotein 1 antibody
- Bone sialoprotein I antibody
see all
Images
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Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)All lanes : Anti-Osteopontin antibody [EPR21139-316] (ab214050) at 1/1000 dilution
Lane 1 : Human urine at 20 µg
Lane 2 : Human fetal kidney lysate at 10 µg
Secondary
Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 35 kDa
Observed band size: 45,60,70,90 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
The multiple bands observed are likely due to expression of different isoforms, as well as glycosylation (PMID: 8761944; PMID: 11375993; PMID: 8535240; PMID: 12890718).
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![Immunoprecipitation - Anti-Osteopontin antibody [EPR21139-316] (ab214050)](https://www.abcam.com/ps/products/214/ab214050/Images/ab214050-311747-anti-osteopontin-antibody-epr21139-316-immunoprecipitation.jpg)
Immunoprecipitation - Anti-Osteopontin antibody [EPR21139-316] (ab214050)Osteopontin was immunoprecipitated from 0.35 mg of human kidney lysate with ab214050 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214050 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution
Lane 1: Human kidney lysate 10 µg (Input).
Lane 2: ab214050 IP in human kidney lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214050 in human kidney lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
The multiple bands observed are likely due to expression of different isoforms, as well as glycosylation (PMID: 8761944; PMID: 11375993; PMID: 8535240; PMID: 12890718).
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![Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)](https://www.abcam.com/ps/products/214/ab214050/Images/ab214050-311745-anti-osteopontin-antibody-epr21139-316-western-blot.jpg)
Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)Anti-Osteopontin antibody [EPR21139-316] (ab214050) at 1/1000 dilution + Human Osteopontin recombinant protein (aa17-300) produced in a mammalian expression at 0.01 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 35 kDa
Observed band size: 60-65 kDa why is the actual band size different from the predicted?
Exposure time: 26 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The his-tagged recombinant protein was produced in a mammalian expression system, thereby allowing it to be post-translationally modified.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21139-316] (ab214050)](https://www.abcam.com/ps/products/214/ab214050/Images/ab214050-311744-anti-osteopontin-antibody-epr21139-316-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21139-316] (ab214050)Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Osteopontin with ab214050 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in bile ducts of human liver (PMID: 16221502). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21139-316] (ab214050)](https://www.abcam.com/ps/products/214/ab214050/Images/ab214050-311743-anti-osteopontin-antibody-epr21139-316-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21139-316] (ab214050)Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Osteopontin with ab214050 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use. Cytoplasmic staining in distal tubules of human kidney (PMID: 17938278), performed on a Leica Biosystems BOND® RX instrument. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat-mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 minutes.
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![Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody [EPR21139-316] (ab214050)](https://www.abcam.com/ps/products/214/ab214050/Images/ab214050-480699-anti-osteopontin-antibody-immunocytochemistry-human.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody [EPR21139-316] (ab214050)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-87 MG cell line labelling Osteopontin with ab214050 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplamic staining in U-87 MG cell line. The nuclear counter stain is DAPI (blue).
The negative controls is as follows:
-ve control: Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
![Anti-Osteopontin antibody [EPR21139-316] (ab214050)](https://www.abcam.com/ps/products/214/ab214050/Images/ab214050-6-benefits-of-recombinant-antibodies.png)
Anti-Osteopontin antibody [EPR21139-316] (ab214050)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (9)
ab214050 has been referenced in 9 publications.
- Zhao L et al. Catalpol Protects Against High Glucose-Induced Bone Loss by Regulating Osteoblast Function. Front Pharmacol 12:626621 (2021). PubMed: 33776769
- Yang L et al. Wnt7a promotes the osteogenic differentiation of human mesenchymal stem cells. Int J Mol Med 47:N/A (2021). PubMed: 33846764
- Li M et al. Calculus-related functional protein expression in ureteral calculus-adhered polyp: A preliminary study. Medicine (Baltimore) 100:e26512 (2021). PubMed: 34160472
- Gao X et al. LINC00899 promotes osteogenic differentiation by targeting miR-374a and RUNX2 expression. Exp Ther Med 22:1071 (2021). PubMed: 34447464
- Xiang J et al. lncRNA SNHG1 attenuates osteogenic differentiation via the miR-101/DKK1 axis in bone marrow mesenchymal stem cells. Mol Med Rep 22:3715-3722 (2020). PubMed: 32901867
- Liu Z et al. Proteomic Analysis of Human Endometrial Tissues Reveals the Roles of PI3K/AKT/mTOR Pathway and Tumor Angiogenesis Molecules in the Pathogenesis of Endometrial Cancer. Biomed Res Int 2020:5273969 (2020). PubMed: 32908897
- Wangzhou K et al. MiR-143-3p Inhibits Osteogenic Differentiation of Human Periodontal Ligament Cells by Targeting KLF5 and Inactivating the Wnt/ß-Catenin Pathway. Front Physiol 11:606967 (2020). PubMed: 33603676
- Zhu L et al. Hyperhomocysteinemia induces vascular calcification by activating the transcription factor RUNX2 via Krüppel-like factor 4 up-regulation in mice. J Biol Chem 294:19465-19474 (2019). PubMed: 31628194
- Chen P et al. Identification and validation of four hub genes involved in the plaque deterioration of atherosclerosis. Aging (Albany NY) 11:6469-6489 (2019). PubMed: 31449494