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Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Osteopontin Recombinant antibody

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

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

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

Conjugate

Unconjugated

CloneNo.

240206A2

Cat no : 83341-1-RR

Synonyms

OPN, SPP1, SPP1, OPN, ETA 1, BSPI

Validation Data Gallery

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  • Western Blot (WB) analysis of various lysates using OPN, SPP1 Recombinant antibody (83341-1-RR)

    WB analysis using 83341-1-RR

    Various lysates were subjected to SDS PAGE followed by western blot with 83341-1-RR (OPN, SPP1 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

  • Immunohistochemistry (IHC) staining of human cervical squamous cancer tissue using Osteopontin Recombinant antibody (83341-1-RR)

    IHC staining of human cervical squamous cancer using 83341-1-RR

    Immunohistochemical analysis of paraffin-embedded human cervical squamous cancer tissue slide using 83341-1-RR (Osteopontin antibody) at dilution of 1:400 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunofluorescence (IF) / fluorescent staining of RAW 264.7 cells using Osteopontin Recombinant antibody (83341-1-RR)

    IF Staining of RAW 264.7 using 83341-1-RR

    Immunofluorescent analysis of (4% PFA) fixed RAW 264.7 cells using OPN, SPP1 antibody (83341-1-RR, Clone: 240206A2 ) at dilution of 1:250 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).

  • Flow cytometry (FC) experiment of U-937 cells using Osteopontin Recombinant antibody (83341-1-RR)

    FC experiment of U-937 using 83341-1-RR

    1x10^6 U-937 cells were intracellularly stained with 0.25 ug OPN, Spp1 Recombinant Antibody (83341-1-RR, Clone:240206A2) and PE-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

  • Flow cytometry (FC) experiment of A549 cells using Osteopontin Recombinant antibody (83341-1-RR)

    FC experiment of A549 using 83341-1-RR

    1x10^6 A549 cells were intracellularly stained with 0.25 ug OPN, Spp1 Recombinant Antibody (83341-1-RR, Clone:240206A2) and PE-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

  • Affinity and Kinetic Characterization of 83341-1-RR

    Affinity and Kinetic Characterization of 83341-1-RR

    Biolayer interferometry (BLl) kinetic assays of 83341-1-RR against Human OPN/SPP1 were performed. The affinity constant is 36.8 nM.

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Tested Applications

Positive WB detected inMOLT-4 cells, HepG2 cells, HEK-293 cells
Positive IHC detected inhuman cervical squamous cancer 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 inRAW 264.7 cells
Positive FC (Intra) detected inU-937 cells, A549 cells

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:1000-1:6000
Immunohistochemistry (IHC)IHC : 1:200-1:800
Immunofluorescence (IF)/ICCIF/ICC : 1:125-1:500
Flow Cytometry (FC) (INTRA)FC (INTRA) : 0.25 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.

Product Information

83341-1-RR targets Osteopontin in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse samples.

Tested Reactivity human, mouse
Host / Isotype Rabbit / IgG
Class Recombinant
Type Antibody
Immunogen Recombinant protein 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 weight60-66 kDa
GenBank accession numberBC007016
Gene symbol Osteopontin
Gene ID (NCBI) 6696
Conjugate Unconjugated
Form Liquid
Purification MethodProtein A purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC 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 83341-1-RRDownload protocol
IHC protocol for Osteopontin antibody 83341-1-RRDownload protocol
IF protocol for Osteopontin antibody 83341-1-RRDownload protocol
Standard Protocols
Click here to view our Standard Protocols