Osteopontin Recombinant antibody, PBS Only (Capture)

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

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

IHC, IF/ICC, Cytometric bead array, Sandwich ELISA, Indirect ELISA, Sample test

Conjugate

Unconjugated

CloneNo.

240206B7

Cat no : 83341-2-PBS

Synonyms

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

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Validation Data Gallery

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  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.

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  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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  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.

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  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.

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  IHC staining of mouse lung using 83341-2-RR (same clone as 83341-2-PBS)

  Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 83341-2-RR (OPN, SPP1 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 83341-2-PBS in a different storage buffer formulation.

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  IHC staining of mouse lung using 83341-2-RR (same clone as 83341-2-PBS)

  Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 83341-2-RR (OPN, SPP1 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 83341-2-PBS in a different storage buffer formulation.

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  IF Staining of HepG2 using 83341-2-RR (same clone as 83341-2-PBS)

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using OPN, SPP1 antibody (83341-2-RR, Clone: 240206B7 ) 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 83341-2-PBS in a different storage buffer formulation.

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  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 83341-2-RR

  Biolayer interferometry (BLl) kinetic assays of 83341-2-RR against Human OPN/SPP1 were performed. The affinity constant is 0.468 nM.

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Product Information

83341-2-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: Fusion Protein
• Full Name: secreted phosphoprotein 1
• Calculated Molecular Weight: 314 aa, 35 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).