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Rabbit Recombinant Monoclonal Osteopontin antibody. Carrier free. Suitable for WB, ICC/IF, IHC-P and reacts with Mouse samples.


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (AB229854), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (AB229854), expandable thumbnail
  • Western blot - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (AB229854), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (AB229854), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IFIHC-P
Mouse
Tested
Tested
Tested

Tested
Tested

Species
Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
-
Notes

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

Associated Products

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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.

Alternative names

Eta-1, Op, Spp-1, Spp1, Osteopontin, 2AR, Bone sialoprotein 1, Calcium oxalate crystal growth inhibitor protein, Early T-lymphocyte activation 1 protein, Minopontin, Secreted phosphoprotein 1, SPP-1

Recommended products

Rabbit Recombinant Monoclonal Osteopontin antibody. Carrier free. Suitable for WB, ICC/IF, IHC-P and reacts with Mouse samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR21138
Purification technique
Affinity purification Protein A
Specificity

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

Osteopontin is a secretory protein, to allow the detection of Osteopontin in cell lysate, we suggest treating cells with BFA which inhibits protein secretion.

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab229854 is the carrier-free version of Anti-Osteopontin antibody [EPR21138] ab218237.

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:

  • - 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 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.

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:
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4 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (ab229854), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (ab229854)

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Osteopontin with Anti-Osteopontin antibody [EPR21138] ab218237 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in distal tubules of mouse kidney (PMID: 17938278). 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Osteopontin antibody [EPR21138] ab218237).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (ab229854), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (ab229854)

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling Osteopontin with Anti-Osteopontin antibody [EPR21138] ab218237 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in biliary epithelial cells of mouse 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Osteopontin antibody [EPR21138] ab218237).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (ab229854), expandable thumbnail

    Western blot - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (ab229854)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM/TBST.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Osteopontin antibody [EPR21138] ab218237).

    All lanes: Western blot - Anti-Osteopontin antibody [EPR21138] (Anti-Osteopontin antibody [EPR21138] ab218237) 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 100 ng/ml lipopolysaccharide (LPS) for 4h whole cell lysate at 20 µg

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

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

  • Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (ab229854), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (ab229854)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labelling Osteopontin with primary antibody anti-Osteopontin (Anti-Osteopontin antibody [EPR21138] ab218237) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody at 1/1000 dilution. Confocal image showing increased cytoplasmic staining in RAW 264.7 cells treated with LPS (100 ng/ml) for 6 h, then add BFA (300 ng/ml) for 3 h. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) was used to counterstain tubulin at 1/200 dilution. The nuclear counter stain is DAPI (blue).
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Osteopontin antibody [EPR21138] ab218237).

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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