Revision 1

#17090

Store at -20C

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IHC-P, IF-F, IF-IC, FC-FP

Reactivity:
M

Sensitivity:
Endogenous

MW (kDa):
58-75

Source/Isotype:
Rabbit IgG

UniProt ID:
#P10923

Entrez-Gene Id:
20750

Product Usage Information

This product is the carrier free version of product #88742. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

BSA and Azide Free antibodies are quality control tested by size exclusion chromatography (SEC) to determine antibody integrity.

Formulation

Supplied in 1X PBS (10 mM Na2HPO4, 3 mM KCl, 2 mM KH2PO4, and 140 mM NaCl (pH 7.8)). BSA and Azide Free.

For standard formulation of this product see product #88742

Storage

Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity/Sensitivity

Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free) recognizes endogenous levels of total osteopontin/SPP1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala149 of mouse osteopontin/SPP1 protein.

Background

Osteopontin is a highly phosphorylated secreted glycoprotein expressed in a variety of cell types. Osteopontin, encoded by the gene secreted phosphoprotein 1 (SPP1), was originally isolated and cloned from a rat osteosarcoma cell line (1). Osteopontin is a highly negatively charged protein that likely regulates extracellular matrix turnover and remodeling, both in bone and other tissue (2). Osteopontin also has three highly conserved receptor-binding domains, otherwise known as arginine-glycine-aspartic acid (RGD) domains. Binding of osteopontin to cell surface integrins likely activates cell signaling pathways that regulate cell proliferation, adhesion, invasion, migration, and fibrosis (3,4). Osteopontin may be regulated by posttranslational regulation, including phosphorylation, glycosylation, and proteolytic cleavage (5). Osteopontin is also linked to several human diseases. For example, cleaved osteopontin is higher in patients suffering with cirrhosis, a disease involving fibrosis of the liver (5). Its role in the brain is unclear, but SPP1 expression in microglia, the resident macrophages of the brain, is upregulated in mouse neurodegenerative disease models, including Alzheimer’s disease (6). Osteopontin’s role in disease likely involves its function as a key mediator of inflammatory responses (4).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

W: Western Blotting IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

M: Mouse

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from RAW 264.7 and Neuro-2a cells, untreated (-) or treated with λ phosphatase (+), using Osteopontin/SPP1 (E9Z1D) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Data were generated using the standard formulation of this product.
Western Blotting Image 1: Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded mouse ovary using Osteopontin/SPP1 (E9Z1D) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 1: Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded mouse pancreas using Osteopontin/SPP1 (E9Z1D) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 2: Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded pons region of mouse brain using Osteopontin/SPP1 (E9Z1D) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 3: Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded renca syngeneic tumor (left), 4T1 syngeneic tumor (middle), or mouse liver (right) using Osteopontin/SPP1 (E9Z1D) Rabbit mAb (top) or an Osteopontin/SPP1 Rabbit mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on mouse Osteopontin/SPP1. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 4: Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded mouse kidney using Osteopontin/SPP1 (E9Z1D) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).  Data were generated using the standard formulation of this product.
Immunohistochemistry Image 5: Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded RAW 264.7 cell pellet (left, positive) or Neuro-2a cell pellet (right, negative) using Osteopontin/SPP1 (E9Z1D) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 6: Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free)
Confocal immunofluorescent analysis of fixed frozen mouse kidney labeled with Osteopontin/SPP1 (E9Z1D) Rabbit mAb (left, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to co-labeling with Ras (E4K9L) Rabbit mAb (Alexa Fluor® 488 Conjugate) #51813 (right, red pseudocolor) and DAPI #4083 (right, blue). Data were generated using the standard formulation of this product.
Immunofluorescence Image 1: Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free)
Confocal immunofluorescent analysis of fixed frozen mouse thalamus labeled with Osteopontin/SPP1 (E9Z1D) Rabbit mAb (left, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to co-labeling with Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #78060 (right, red) and DAPI #4083 (right, blue). Data were generated using the standard formulation of this product.
Immunofluorescence Image 2: Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of fixed frozen mouse pons labeled with Osteopontin/SPP1 (E4O2F) Rabbit mAb (left, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to colabeling with Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #78060 (right, red) and DAPI #4083 (right, blue). Data were generated using the standard formulation of this product.
Immunofluorescence Image 3: Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free)
Confocal immunofluorescent analysis of RAW 264.7 cells (left, positive) and Neuro-2a cells (right, negative) using Osteopontin/SPP1 (E9Z1D) Rabbit mAb (green) and DAPI #4083 (blue). Cells were treated with Brefeldin A #9972 (100 ng/mL, 90 min) to inhibit protein secretion. Data were generated using the standard formulation of this product.
Immunofluorescence Image 1: Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free)
Flow cytometric analysis of Neuro-2a cells (blue, negative) and RAW 264.7 cells (green, positive) using Osteopontin/SPP1 (E9Z1D) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Data were generated using the standard formulation of this product.
Flow Cytometry Image 1: Osteopontin/SPP1 (E9Z1D) Rabbit Monoclonal Antibody (BSA and Azide Free)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.