Osteopontin Recombinant antibody

Osteopontin Uni-rAb^TM 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

• 

  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.

• 

  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).

• 

  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).

• 

  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.

---

Tested Applications

• Positive WB detected in: MOLT-4 cells, HepG2 cells, HEK-293 cells
• Positive IHC detected in: human 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 in: RAW 264.7 cells
• Positive FC (Intra) detected in: U-937 cells, A549 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• Immunohistochemistry (IHC): IHC : 1:200-1:800
• Immunofluorescence (IF)/ICC: IF/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
• Full Name: secreted phosphoprotein 1
• Calculated Molecular Weight: 314 aa, 35 kDa
• Observed Molecular Weight: 60-66 kDa
• GenBank Accession Number: BC007016
• Gene Symbol: Osteopontin
• Gene ID (NCBI): 6696
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A 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).