Osteopontin Recombinant antibody

Osteopontin Uni-rAb^TM Recombinant Antibody for WB, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

2

CloneNo.

6B11

Cat no : 80912-4-RR

Synonyms

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

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Validation Data Gallery

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  WB analysis using 80912-4-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 80912-4-RR (OPN, SPP1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  FC experiment of HepG2 using 80912-4-RR

  1x10^6 HepG2 cells were intracellularly stained with 0.25 ug OPN, SPP1 Recombinant antibody (80912-4-RR, Clone:6B11) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2)(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 80912-4-RR

  Biolayer interferometry (BLl) kinetic assays of 80912-4-RR against Human OPN, SPP1 were performed. The affinity constant is 199 nM.

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Tested Applications

• Positive WB detected in: HEK-293 cells, HepG2 cells, C2C12 cells, mouse kidney tissue, rat kidney tissue
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:12800
• 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.

Published Applications

• WB: See 2 publications below

Product Information

80912-4-RR targets Osteopontin in WB, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: Osteopontin fusion protein Ag19216
• Full Name: secreted phosphoprotein 1
• Calculated Molecular Weight: 314 aa, 35 kDa
• Observed Molecular Weight: 70 kDa, 44-66 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).

Publications

Species	Application	Title
human	WB	Cell Death Dis; LINC01133 promotes pancreatic ductal adenocarcinoma epithelial-mesenchymal transition mediated by SPP1 through binding to Arp3; Authors - Yefan Yang
human	WB	J Neurooncol; Exploring the prognostic value of BRMS1 + microglia based on single-cell anoikis regulator patterns in the immunologic microenvironment of GBM; Authors - Songyun Zhao