Osteopontin Polyclonal antibody

• Featured Product
• KD/KO Validated

Osteopontin Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (2)

Applications

WB, IHC, IF/ICC, FC (Intra), CoIP, ELISA

Conjugate

Unconjugated

Publications

341

Reviews (9)

Cat no : 22952-1-AP

Synonyms

SPP1, PSEC0156, OPN, ETA 1, BSPI

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Validation Data Gallery

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  WB analysis using 22952-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 22952-1-AP (Osteopontin antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of Jurkat using 22952-1-AP

  Jurkat cells were subjected to SDS PAGE followed by western blot with 22952-1-AP (Osteopontin antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HEK-293 using 22952-1-AP

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 22952-1-AP (Osteopontin antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HepG2 using 22952-1-AP

  HepG2 cells were subjected to SDS PAGE followed by western blot with 22952-1-AP (Osteopontin antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 22952-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 22952-1-AP (Osteopontin antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.

• 

  IHC staining of mouse kidney using 22952-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 22952-1-AP (Osteopontin antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse kidney using 22952-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 22952-1-AP (Osteopontin antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse lung using 22952-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 22952-1-AP (Osteopontin antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse lung using 22952-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 22952-1-AP (Osteopontin antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HepG2 using 22952-1-AP

  Immunofluorescent analysis of HepG2 cells using 22952-1-AP (Osteopontin antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of HEK-293 using 22952-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HEK-293 cells using Osteopontin antibody (22952-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

• 

  IF Staining of HepG2 using 22952-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using Osteopontin antibody (22952-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

• 

  FC experiment of HepG2 using 22952-1-AP

  1X10^6 HepG2 cells were intracellularly stained with 0.4 ug Anti-Human Osteopontin (22952-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP, Clone: ) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: HEK-293 cells, Jurkat cells, C2C12 cell, HepG2 cells, mouse kidney tissue
• Positive IHC detected in: mouse kidney tissue, mouse lung tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HepG2 cells, HEK-293 cells
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:4000
• Immunohistochemistry (IHC): IHC : 1:250-1:1000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 9 publications below
• WB: See 228 publications below
• IHC: See 82 publications below
• IF: See 88 publications below
• ELISA: See 1 publications below
• CoIP: See 2 publications below

Product Information

22952-1-AP targets Osteopontin in WB, IHC, IF/ICC, FC (Intra), CoIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, rabbit, bovine
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: Osteopontin fusion protein Ag19216
• Full Name: secreted phosphoprotein 1
• Calculated Molecular Weight: 314 aa, 35 kDa
• Observed Molecular Weight: 70 kDa, 44-66 kDa
• GenBank Accession Number: BC007016
• Gene Symbol: Osteopontin
• Gene ID (NCBI): 6696
• RRID: AB_2783651
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Osteopontin (OPN), also known as SPP1, is a secreted glycophosphoprotein that belongs to the small integrin-binding ligand N-linked glycoprotein (SIBLING) family. Originally isolated from bone, OPN has been found in kidneys, vascular tissues, biological fluids, and various tumor tissues (PMID: 15138464; 16406521). OPN can interact with CD44 and integrins and regulate diverse biological processes. It has a multifaceted role in bone development and remodeling, and is also involved in the inflammatory and immune response, oncogenesis and cancer progression. The very acidic nature of OPN, as well as the presence of variable posttranslational modifications, has led to anomalous migration in SDS-polyacrylamide gels and therefore to reports of different molecular weights for OPN (PMID: 8293561). Depending on the cell and tissue source and/or the SDS-PAGE system, OPN migrates with a molecular weight of 44-80 kDa, as well as at some smaller bands corresponding to peptide fragments (PMID: 8195113; 17890765).

Publications

Species	Application	Title
mouse	WB	Circulation; Carbonylation of Runx2 at K176 by 4-Hydroxynonenal Accelerates Vascular Calcification; Authors - Xiaoxuan Zhai
mouse	IF	Adv Mater; Realizing Highly Efficient Sonodynamic Bactericidal Capability through the Phonon-Electron Coupling Effect Using Two-Dimensional Catalytic Planar Defects; Authors - Congyang Mao
human	IF	Cell Stem Cell; Decoding the temporal and regional specification of microglia in the developing human brain.; Authors - Yanxin Li
rat,human	IF	Bioact Mater; Urchin-like multiscale structured fluorinated hydroxyapatite as versatile filler for caries restoration dental resin composites; Authors - Shuning Zhang
human	IF	Nat Cell Biol; Single-cell reconstruction of the adult human heart during heart failure and recovery reveals the cellular landscape underlying cardiac function.; Authors - Li Wang
mouse	IF	Cell Host Microbe; Gut microbiome dysbiosis contributes to abdominal aortic aneurysm by promoting neutrophil extracellular trap formation; Authors - Zhenyu Tian

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Shinford (Verified Customer) (10-26-2023)

The WB bands exhibit remarkable strength and clarity, with various treatments leading to significant changes in expression.

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Lung cancer cell lines and lung cancer tissues with different treatment

Osteopontin Antibody Western Blot validation (1:1000 dilution) in Lung cancer cell lines and lung cancer tissues with different treatment (Cat no:22952-1-AP)

FH

Shinford (Verified Customer) (09-24-2023)

This antibody works well in esophageal cancer and lung cancer cell lines, and the detection antibody band is clear. It is recommended to use a diluted concentration of 1:2000.

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Cancer cell lines

Osteopontin Antibody Western Blot validation (1:1000 dilution) in Cancer cell lines (Cat no:22952-1-AP)

FH

Elisa (Verified Customer) (08-16-2023)

Tested on mouse skull sections, with appropriate blocking: give a nice stain of osteopontin, but maybe it can be optimized to be able to use an higher dilution.

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:100
• Cell Tissue Type: Mouse skull

FH

Davide (Verified Customer) (09-20-2022)

The antibody works really well. Fantastic product, and the possibility to try the product with the free sample is something really appreciated. Will surely buy a full vial soon.

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:100
• Cell Tissue Type: Cholangiocytes

Osteopontin Antibody Immunofluorescence validation (1:100 dilution) in Cholangiocytes (Cat no:22952-1-AP)

FH

WEI (Verified Customer) (03-08-2022)

Strong specific Bands

• Applications: Western Blot
• Primary Antibody Dilution: 1:3000
• Cell Tissue Type: Mouse Heart

FH

LUNFENG (Verified Customer) (01-27-2020)

VERY GOOD

• Applications: Immunofluorescence,
• Cell Tissue Type: HEART

FH

Jie (Verified Customer) (01-23-2020)

Works with one specific band and one faint non-specific band. However, can be distinguished by protein size. So works fine in general.

• Applications: Western Blot, Immunohistochemistry, Immunofluorescence,
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Mouse kidney

FH

Jingwen (Verified Customer) (01-17-2020)

This antibody showed the specific band and staining results in both western blot and immunofluorescence staining. I strongly recommend it.

• Applications: Western Blot, Immunofluorescence,
• Primary Antibody Dilution: 1:1000 for WB, 1:100 for IF
• Cell Tissue Type: Mouse heart and cultured cardiomyocytes.

FH

Barbara (Verified Customer) (09-28-2019)

Fluorescence-based detection of OPN (green) expression in paraffin-embedded human aortic tissue, heat mediated antigen retrival with 10mM citrate buffer (pH 6,0)

• Applications: Immunohistochemistry,
• Primary Antibody Dilution: 1:150
• Cell Tissue Type: aorta

Osteopontin Antibody Immunohistochemistry, validation (1:150 dilution) in aorta (Cat no:22952-1-AP)

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