Osteopontin Recombinant antibody, PBS Only (Detector)

Osteopontin Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), Cytometric bead array, Sandwich ELISA, Indirect ELISA, Sample test

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IHC, IF/ICC, FC (Intra), Cytometric bead array, Sandwich ELISA, Indirect ELISA, Sample test

Conjugate

Unconjugated

CloneNo.

240206A2

Cat no : 83341-1-PBS

Synonyms

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

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Validation Data Gallery

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  WB analysis using 83341-1-RR (same clone as 83341-1-PBS)

  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. This data was developed using the same antibody clone with 83341-1-PBS in a different storage buffer formulation.

• 

  Sandwich ELISA standard curve of MP00375-1

  Sandwich ELISA standard curve of MP00375-1, Human OPN, SPP1 Recombinant Matched Antibody Pair - PBS only. 83341-2-PBS was coated to a plate as the capture antibody and incubated with serial dilutions of standard Eg0754. 83341-1-PBS was HRP conjugated as the detection antibody. Range: 0.313-20 ng/mL.

• 

  Sample test of MP00375-1

  Urine of eight individual healthy human donors was measured. The Osteopontin/OPN concentration of detected samples was determined to be 1,426.23 ng/mL with a range of 136.64 - 5,219.95 ng/mL.

• 

  Sample test of MP00375-1

  Milk of seven individual healthy human donors was measured. The Osteopontin/OPN concentration of detected samples was determined to be 35,304.3 ng/mL with a range of 13,338.9 - 63,338.0 ng/mL.

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  Sample test of MP00375-1

  HepG2 (human hepatocellular carcinoma cells) were cultured in DMEM supplemented with 10% fetal bovine serum, 2.5 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. An aliquot of the cell culture supernate was removed, assayed for human Osteopontin/OPN, and measured 240.9 ng/mL. A431 were cultured in DMEM supplemented with 10% fetal bovine serum, 2.5 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. An aliquot of the cell culture supernate was removed, assayed for human Osteopontin/OPN, and measured 0.25 ng/mL.

• 

  Sample test of MP00375-1

  Plasma of sixteen individual healthy human donors was measured. The Osteopontin/OPN concentration of detected samples was determined to be 40.58 ng/mL with a range of 7.07 - 92.57 ng/mL.

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  IHC staining of human cervical squamous cancer using 83341-1-RR (same clone as 83341-1-PBS)

  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). This data was developed using the same antibody clone with 83341-1-PBS in a different storage buffer formulation.

• 

  IF Staining of RAW 264.7 using 83341-1-RR (same clone as 83341-1-PBS)

  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). This data was developed using the same antibody clone with 83341-1-PBS in a different storage buffer formulation.

• 

  FC experiment of U-937 using 83341-1-RR (same clone as 83341-1-PBS)

  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). This data was developed using the same antibody clone with 83341-1-PBS in a different storage buffer formulation.

• 

  FC experiment of A549 using 83341-1-RR (same clone as 83341-1-PBS)

  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). This data was developed using the same antibody clone with 83341-1-PBS in a different storage buffer formulation.

• 

  Cytometric bead array standard curve of MP00375-1

  Cytometric bead array standard curve of MP00375-1, OPN, SPP1 Recombinant Matched Antibody Pair, PBS Only. Capture antibody: 83341-2-PBS. Detection antibody: 83341-1-PBS. Standard: Eg0754. Range: 0.313-40 ng/mL.

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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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Product Information

83341-1-PBS targets Osteopontin as part of a matched antibody pair:

MP00375-1: 83341-2-PBS capture and 83341-1-PBS detection (validated in Cytometric bead array, Sandwich ELISA)

Unconjugated rabbit recombinant monoclonal antibody in PBS only (BSA and azide free) storage buffer at a concentration of 1 mg/mL, ready for conjugation. Created using Proteintech’s proprietary in-house recombinant technology. Recombinant production enables unrivalled batch-to-batch consistency, easy scale-up, and future security of supply.

This conjugation ready format makes antibodies ideal for use in many applications including: ELISAs, multiplex assays requiring matched pairs, mass cytometry, and multiplex imaging applications.Antibody use should be optimized by the end user for each application and assay.

• 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 Only
• Storage Conditions: Store at -80°C.

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).