Osteopontin Rekombinanter Antikörper

Osteopontin Rekombinant Antikörper für WB, IHC, IF/ICC, FC (Intra), ELISA

Wirt / Isotyp

Kaninchen / IgG

Getestete Reaktivität

human, Maus

Anwendung

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

Konjugation

Unkonjugiert

Publikationen(2)

CloneNo.

240206A2

Kat-Nr. : 83341-1-RR

Synonyme

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

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  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.

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  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).

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  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).

• 

  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).

• 

  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).

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  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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Geprüfte Anwendungen

• Erfolgreiche Detektion in WB: MOLT-4-Zellen, HEK-293-Zellen, HepG2-Zellen
• Erfolgreiche Detektion in IHC: human cervical squamous cancer tissue; Hinweis: Antigendemaskierung mit TE-Puffer pH 9,0 empfohlen. (*) Wahlweise kann die Antigendemaskierung auch mit Citratpuffer pH 6,0 erfolgen.
• Erfolgreiche Detektion in IF/ICC: RAW 264.7-Zellen
• Erfolgreiche Detektion in FC (Intra): U-937-Zellen, A549-Zellen

Empfohlene Verdünnung

• Western Blot (WB): WB : 1:1000-1:6000
• Immunhistochemie (IHC): IHC : 1:200-1:800
• Immunfluoreszenz (IF)/ICC: IF/ICC : 1:125-1:500
• Durchflusszytometrie (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

Veröffentlichte Anwendungen

• WB: See 1 publications below
• IF: See 2 publications below

Produktinformation

83341-1-RR bindet in WB, IHC, IF/ICC, FC (Intra), ELISA Osteopontin und zeigt Reaktivität mit human, Maus

• Getestete Reaktivität: human, Maus
• In Publikationen genannte Reaktivität: human, Maus
• Wirt / Isotyp: Kaninchen / IgG
• Klonalität: Rekombinant
• Typ: Antikörper
• Immunogen: Rekombinantes Protein
• Vollständiger Name: secreted phosphoprotein 1
• Berechnetes Molekulargewicht: 314 aa, 35 kDa
• Beobachtetes Molekulargewicht: 60-66 kDa
• GenBank-Zugangsnummer: BC007016
• Gene symbol: Osteopontin
• Gene ID (NCBI): 6696
• Konjugation: Unkonjugiert
• Form: Liquid
• Reinigungsmethode: Protein-A-Reinigung
• Lagerungspuffer: PBS with 0.02% sodium azide and 50% glycerol
• Lagerungsbedingungen: Bei -20°C lagern. Nach dem Versand ein Jahr lang stabil Aliquotieren ist bei -20^oC Lagerung nicht notwendig. 20ul Größen enthalten 0,1% BSA.

Hintergrundinformationen

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).

Publikationen

Species	Application	Title
human	IF	J Gastroenterol; Single-cell RNA sequencing and functional analysis reveal the role of altered glycosylation levels of hepatic macrophages in liver cirrhosis; Authors - Chunmei Wen
mouse	WB,IF	Tissue Cell; GIT2 negatively regulates the NF-κB pathway directly or indirectly by regulating TRAF3 expression to promote osteogenic differentiation of BMSCs; Authors - Yanna Wang