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Rabbit Recombinant Multiclonal Osteopontin antibody. Suitable for IP, WB, I-ELISA, IHC-P and reacts with Mouse, Human, Recombinant fragment - Human, Recombinant fragment - Mouse samples. Cited in 3 publications.

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Images

Western blot - Anti-Osteopontin antibody [RM1018] (AB283656), expandable thumbnail
  • Western blot - Anti-Osteopontin antibody [RM1018] (AB283656), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [RM1018] (AB283656), expandable thumbnail
  • Immunoprecipitation - Anti-Osteopontin antibody [RM1018] (AB283656), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [RM1018] (AB283656), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Multiclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBI-ELISAICC/IFFlow Cyt (Intra)IHC-FrIHC-P
Human
Tested
Tested
Expected
Not recommended
Not recommended
Not recommended
Tested
Mouse
Tested
Tested
Expected
Not recommended
Not recommended
Not recommended
Tested
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Recombinant fragment - Human
Not recommended
Not recommended
Tested
Not recommended
Not recommended
Not recommended
Not recommended
Recombinant fragment - Mouse
Not recommended
Not recommended
Tested
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Mouse
Dilution info
1/30
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Human
Dilution info
1/30
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Rat, Recombinant fragment - Human, Recombinant fragment - Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/1000
Notes

-

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Rat, Recombinant fragment - Human, Recombinant fragment - Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Recombinant fragment - Human
Dilution info
0.06 µg/mL
Notes

-

Species
Recombinant fragment - Mouse
Dilution info
0.06 µg/mL
Notes

-

Expected
Expected

Species
Mouse, Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Human, Rat, Recombinant fragment - Human, Recombinant fragment - Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Human, Rat, Recombinant fragment - Human, Recombinant fragment - Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Rat, Mouse, Human, Recombinant fragment - Human, Recombinant fragment - Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/2000
Notes

-

Species
Human
Dilution info
1/2000
Notes

-

Not recommended
Not recommended

Species
Rat, Recombinant fragment - Human, Recombinant fragment - Mouse
Dilution info
-
Notes

-

Target data

Function

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.

Additional Targets

Spp1

Alternative names

BNSP, OPN, PSEC0156, SPP1, Osteopontin, Bone sialoprotein 1, Nephropontin, Secreted phosphoprotein 1, Urinary stone protein, Uropontin, SPP-1

Recommended products

Rabbit Recombinant Multiclonal Osteopontin antibody. Suitable for IP, WB, I-ELISA, IHC-P and reacts with Mouse, Human, Recombinant fragment - Human, Recombinant fragment - Mouse samples. Cited in 3 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Multiclonal
Immunogens
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
RM1018
Purification technique
Affinity purification Protein A
Specificity

This antibody is not recommended for use in WB with mouse tissue samples.

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

This product is a recombinant multiclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Western blot - Anti-Osteopontin antibody [RM1018] (ab283656), expandable thumbnail

    Western blot - Anti-Osteopontin antibody [RM1018] (ab283656)

    Blocking and diluting buffer and concentration: 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).

    Exposure time: 2 min.

    All lanes: Western blot - Anti-Osteopontin antibody [RM1018] (ab283656) at 1/1000 dilution

    Lane 1: Human kidney lysate at 20 µg

    Lane 2: MDA-MB-435S (Human mammary gland ductal carcinoma melanocyte) whole cell lysate at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 35 kDa

    Observed band size: 60 kDa

  • Western blot - Anti-Osteopontin antibody [RM1018] (ab283656), expandable thumbnail

    Western blot - Anti-Osteopontin antibody [RM1018] (ab283656)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Exposure time: 37 seconds.

    All lanes: Western blot - Anti-Osteopontin antibody [RM1018] (ab283656) at 1/1000 dilution

    Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

    Lane 2: RAW 264.7 treated with 100ng/ml lipopolysaccharide (LPS) for 3h then add 300ng/ml BFA for another 3h whole cell lysate at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 35 kDa

    Observed band size: 60 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [RM1018] (ab283656), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [RM1018] (ab283656)

    Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling Osteopontin with ab283656 at 1/2000 (0.232 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the bile ducts in human liver. The section was incubated with ab283656 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunoprecipitation - Anti-Osteopontin antibody [RM1018] (ab283656), expandable thumbnail

    Immunoprecipitation - Anti-Osteopontin antibody [RM1018] (ab283656)

    Osteopontin was immunoprecipitated from 0.35 mg Human kidney lysate 10ug with ab283656 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283656 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: Human kidney lysate 10ug

    Lane 2: ab283656 IP in Human kidney lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab283656 in Human kidney lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 min.

    All lanes: Immunoprecipitation - Anti-Osteopontin antibody [RM1018] (ab283656)

    Predicted band size: 35 kDa

    Observed band size: 60 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [RM1018] (ab283656), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [RM1018] (ab283656)

    Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling Osteopontin with ab283656 at 1/2000 (0.232 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on distal tubules in human kidney. The section was incubated with ab283656 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

  • Immunoprecipitation - Anti-Osteopontin antibody [RM1018] (ab283656), expandable thumbnail

    Immunoprecipitation - Anti-Osteopontin antibody [RM1018] (ab283656)

    Osteopontin was immunoprecipitated from 0.35 mg RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 3h then add 300ng/ml BFA for another 3h whole cell lysate 10ug with ab283656 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283656 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 3h then add 300ng/ml BFA for another 3h whole cell lysate 10ug

    Lane 2: ab283656 IP in RAW 264.7 treated with 100ng/ml LPS for 3h then add 300ng/ml BFA for another 3h whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab283656 in RAW 264.7 treated with 100ng/ml LPS for 3h then add 300ng/ml BFA for another 3h whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 32 seconds.

    Lower bands should be MMP-cleaved Osteopontin. (PMID: 11375993)

    All lanes: Immunoprecipitation - Anti-Osteopontin antibody [RM1018] (ab283656)

    Predicted band size: 35 kDa

    Observed band size: 60 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [RM1018] (ab283656), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [RM1018] (ab283656)

    Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labelling Osteopontin with ab283656 at 1/2000 (0.232 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the bile ducts in mouse liver. The section was incubated with ab283656 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Competitive ELISA - Anti-Osteopontin antibody [RM1018] (ab283656), expandable thumbnail

    Competitive ELISA - Anti-Osteopontin antibody [RM1018] (ab283656)

    ELISA using ab283656 at varying antibody concentrations and antigen concentration at 1000 ng/ml. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

  • Western blot - Anti-Osteopontin antibody [RM1018] (ab283656), expandable thumbnail

    Western blot - Anti-Osteopontin antibody [RM1018] (ab283656)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as a GAPDH loading control.

    This antibody is not recommended for use in WB with mouse tissue samples.
    We suggest treating cells with BFA which inhibits protein secretion.

    All lanes: Western blot - Anti-Osteopontin antibody [RM1018] (ab283656) at 1/1000 dilution

    Lane 1: Mouse kidney tissue lysate at 20 µg

    Lane 2: Mouse placenta tissue lysate at 20 µg

    Lane 3: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

    Lane 4: RAW 264.7 treated with 100ng/ml lipopolysaccharide (LPS) for 4h whole cell lysate at 20 µg

    Lane 5: RAW 264.7 treated with 1000ng/ml BFA for 3h whole cell lysate at 20 µg

    Lane 6: RAW 264.7 treated with 100ng/ml lipopolysaccharide (LPS) for 4h then add 1000ng/ml BFA for another 3h whole cell lysate at 20 µg

    Lane 7: Human kidney tissue lysate at 20 µg

    Lane 8: A549 (Human lung carcinoma epithelial cell) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

    Predicted band size: 35 kDa

    Observed band size: 60 kDa

    Exposure time: 180s

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Product protocols

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