Rabbit Recombinant Monoclonal Osteopontin antibody. Carrier free. Suitable for ICC/IF, IHC-P, IP, WB and reacts with Human, Recombinant fragment samples.
pH: 7.2 - 7.4
Constituents: PBS
ICC/IF | IHC-P | IP | WB | |
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Human | Tested | Tested | Tested | Expected |
Recombinant fragment | Not recommended | Not recommended | Not recommended | Expected |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Recombinant fragment | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Recombinant fragment | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Recombinant fragment | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Recombinant fragment, Human | Dilution info Use at an assay dependent concentration. | Notes - |
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Major non-collagenous bone protein that 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.
BNSP, OPN, PSEC0156, SPP1, Osteopontin, Bone sialoprotein 1, Nephropontin, Secreted phosphoprotein 1, Urinary stone protein, Uropontin, SPP-1
Rabbit Recombinant Monoclonal Osteopontin antibody. Carrier free. Suitable for ICC/IF, IHC-P, IP, WB and reacts with Human, Recombinant fragment samples.
pH: 7.2 - 7.4
Constituents: PBS
Osteopontin is a secretory protein, to allow the detection of Osteopontin in cell lysate, we suggest treating cells with BFA which inhibits protein secretion.
ab236213 is the carrier-free version of Anti-Osteopontin antibody [EPR21139-316] ab214050.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Osteopontin also known as OPN or secreted phosphoprotein 1 (SPP1) is a glycoprotein with a molecular weight of approximately 44 to 75 kDa depending on its post-translational modifications. Osteopontin is characteristically expressed in bone but is also present in various body tissues including kidney liver and immune system cells. This multifunctional protein interacts with cell surface receptors such as integrins and CD44 playing an important role in cell signaling and extracellular matrix interactions. Researchers often study it using osteopontin western blot or osteopontin ELISA kit to understand its expression patterns and roles.
Osteopontin affects cell adhesion migration and survival. It does not form part of a known large complex but interacts closely with its integrin receptors. Osteopontin modulates immune responses and aids in bone remodeling by affecting osteoclast and osteoblast functions. It is also influential in inflammation processes where it orchestrates the recruitment and activation of macrophages. The osteopontin function is significant in both physiological processes and pathologies marking it as an important protein to study in numerous biological contexts.
Osteopontin is integral to both the bone remodeling and immune response pathways. It collaborates with proteins like osteocalcin and matrix metalloproteinases within these processes. In the bone remodeling pathway osteopontin mediates communication between osteoclasts and osteoblasts impacting bone resorption and formation. In the immune response pathway it coordinates with cytokines such as interleukin-6 to modulate immune cell activities. Its roles in these pathways indicate its multifunctional nature and its impact across different biological systems.
Osteopontin links to cancer and cardiovascular diseases. It facilitates tumor progression and metastasis by interacting with proteins such as vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9). In cardiovascular diseases osteopontin influences atherosclerosis development through interactions with inflammatory cytokines and contributes to plaque stability. These disease associations highlight the importance of osteopontin as both a therapeutic target and a biomarker for disease progression and prognosis.
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We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Osteopontin was immunoprecipitated from 0.35 mg of human kidney lysate with Anti-Osteopontin antibody [EPR21139-316] ab214050 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-Osteopontin antibody [EPR21139-316] ab214050 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5000 dilution.
Lane 1: Human kidney lysate 10 μg (Input).
Lane 2: Anti-Osteopontin antibody [EPR21139-316] ab214050 IP in human kidney lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Osteopontin antibody [EPR21139-316] 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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Osteopontin antibody [EPR21139-316] ab214050).
All lanes: Immunoprecipitation - Anti-Osteopontin antibody [EPR21139-316] (Anti-Osteopontin antibody [EPR21139-316] ab214050)
Predicted band size: 35 kDa
Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Osteopontin with Anti-Osteopontin antibody [EPR21139-316] 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Osteopontin antibody [EPR21139-316] ab214050).
Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Osteopontin with Anti-Osteopontin antibody [EPR21139-316] ab214050 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Osteopontin antibody [EPR21139-316] ab214050).
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).
The identity of the 90kDa band is unknown.
This data was developed using Anti-Osteopontin antibody [EPR21139-316] ab214050, the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide.
All lanes: Western blot - Anti-Osteopontin antibody [EPR21139-316] (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
Lane 1: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution
Lane 2: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 35 kDa
Observed band size: 45 kDa, 60 kDa, 70 kDa
Exposure time: 26s
Blocking and diluting buffer concentration: 5% NFDM/TBST.
We suggest treating cells with BFA which inhibits protein secretion.
This data was developed using Anti-Osteopontin antibody [EPR21139-316] ab214050, the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide.
All lanes: Western blot - Anti-Osteopontin antibody [EPR21139-316] (Anti-Osteopontin antibody [EPR21139-316] ab214050) at 1/1000 dilution
Lane 1: Untreated HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: HepG2 treated with 300ng/ml BFA for 3h whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 35 kDa
Observed band size: 60 kDa
Exposure time: 60s
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