Validation Data Gallery

Filter: AllWBIHCIF/ICCFC (Intra)

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 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.
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.
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).
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).
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).

Featured Product
KD/KO Validated

Osteopontin Polyclonal antibody

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

Cat No. 22952-1-AP

Publications(361)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (2)

Applications

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

Synonyms

SPP1, PSEC0156, OPN, ETA 1, BSPI

Formulation: PBS and Azide

PBS and Azide

Conjugate: Unconjugated

Unconjugated

CoraLite® Plus 488 CoraLite® Plus 647

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (361)
Protocols

データシート［PDF］を印刷

MSDS

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

Application	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 240 publications below
IHC	See 86 publications below
IF	See 93 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 相同性解析による交差性が予測される生物種
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
UNIPROT ID	P10451
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.

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).

Protocols

Product Specific Protocols
WB protocol for Osteopontin antibody 22952-1-AP	Download protocol
IHC protocol for Osteopontin antibody 22952-1-AP	Download protocol
IF protocol for Osteopontin antibody 22952-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
mouse	WB	Circulation Carbonylation of Runx2 at K176 by 4-Hydroxynonenal Accelerates Vascular Calcification Authors - Xiaoxuan Zhai View Article
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 View Article
human	IF	Cell Stem Cell Decoding the temporal and regional specification of microglia in the developing human brain. Authors - Yanxin Li View Article
rat,human	IF	Bioact Mater Urchin-like multiscale structured fluorinated hydroxyapatite as versatile filler for caries restoration dental resin composites Authors - Shuning Zhang View Article
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 View Article
mouse	IF	Cell Host Microbe Gut microbiome dysbiosis contributes to abdominal aortic aneurysm by promoting neutrophil extracellular trap formation Authors - Zhenyu Tian View Article

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis using 22952-1-AP

WB analysis of Jurkat using 22952-1-AP

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

WB analysis using 22952-1-AP

WB analysis of HepG2 using 22952-1-AP

IHC staining of mouse lung using 22952-1-AP

IHC staining of mouse lung using 22952-1-AP

IHC staining of mouse kidney using 22952-1-AP

IHC staining of mouse kidney using 22952-1-AP

IF Staining of HepG2 using 22952-1-AP

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

IF Staining of HepG2 using 22952-1-AP

FC experiment of HepG2 using 22952-1-AP

Osteopontin Polyclonal antibody

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

-/ -

Tested Applications

Recommended dilution

Published Applications

Product Information

Background Information

Protocols

Publications

Carbonylation of Runx2 at K176 by 4-Hydroxynonenal Accelerates Vascular Calcification

Realizing Highly Efficient Sonodynamic Bactericidal Capability through the Phonon-Electron Coupling Effect Using Two-Dimensional Catalytic Planar Defects

Decoding the temporal and regional specification of microglia in the developing human brain.

Urchin-like multiscale structured fluorinated hydroxyapatite as versatile filler for caries restoration dental resin composites

Single-cell reconstruction of the adult human heart during heart failure and recovery reveals the cellular landscape underlying cardiac function.

Gut microbiome dysbiosis contributes to abdominal aortic aneurysm by promoting neutrophil extracellular trap formation