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Human CCN3 / NOV / IGFBP9 ELISA Pair Set

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Materials provided
Capture Ab:0.5 mg/mL of rabbit anti-NOV monoclonal antibody, Dilute to a working concentration of 2.0 μg/mL in CBS before coating.
Detection Ab:0.5 mg/mL mouse anti-NOV monoclonal antibody conjugated to horseradish-peroxidase (HRP). Dilute to working concentration of 0.05 μg/mL in detection antibody dilution buffer before use.
Standard:Each vial contains 100 ng of recombinant NOV. Reconstitute standard powder with 1 mL detection antibody dilution buffer. After reconstitution, store at -20℃ to -70℃ in a manual defrost freezer. A seven-point standard curve usi ng 2-fold serial dilutions in sample dilution buffer, and a high standard of 2000 pg/mL is recommended.
Sensitivity
The minimum detectable dose of Human CCN3 / NOV / IGFBP9 was determined to be approximately 31.25 pg/ml. This is defined as at least three times standard deviations above the mean optical density of 10 replicates of the zero standard.
Principle of the product
The Human CCN3 / NOV / IGFBP9 ELISA Pair Set is for the quantitative determination of Human CCN3 / NOV / IGFBP9.
This ELISA Pair Set contains the basic components required for the development of sandwich ELISAs.
The Sino Biological ELISA Pair Set is a solid phase sandwich ELISA (Enzyme-Linked Immunosorbent Assay). It utilizes a monoclonal antibody specific for CCN3 / NOV / IGFBP9 coated on a 96-well plate. Standards and samples are added to the wells, and any CCN3 / NOV / IGFBP9 present binds to the immobilized antibody. The wells are washed and a horseradish peroxidase conjugated mouse anti-CCN3 / NOV / IGFBP9 monoclonal antibody is then added, producing an antibody-antigen-antibody "sandwich". The wells are again washed and TMB substrate solution is loaded, which produces color in proportion to the amount of CCN3 / NOV / IGFBP9 present in the sample. To end the enzyme reaction, the stop solution is added and absorbances of the microwell are read at 450 nm.
Storage
Capture Antibody: Aliquot and store at -20℃ to -70℃ for up to 6 months from date of receipt. Avoid repeated freeze-thaw cycles.
Detection Antibody: Protect it from prolonged exposure to light. Aliquot and store at -20℃ to -70℃ and for up to 6 months from date of receipt. Avoid repeated freeze-thaw cycles.
Standard: Store lyophilized Standard at -20℃ to -70℃ for up to 6 months from date of receipt. Aliquot and store the reconstituted Standard at -20℃ to -70℃ for up to 1 month. Avoid repeated freeze-thaw cycles.
Background

Protein NOV homolog, also known as Nephroblastoma-overexpressed gene protein homolog, NOV, and CCN3, is a putative ligand for integrin receptors, is tightly associated with the extracellular matrix and mediates diverse cellular functions, including cell adhesion and proliferation. CCN3 has been shown to negatively regulate growth although it promotes migration in a cell type-specific manner. This secreted protein belongs to the CCN family, and its expression was observed in a broad variety of tissues from the early stage of development , and altered expression of CCN3 has been observed in a variety of tumors, including hepatocellular carcinomas, Wilm's tumors, Ewing's sarcomas, gliomas, rhabdomyosarcomas, and adrenocortical carcinomas. Mature CCN3 protein has five distinct modules and truncated protein variants with altered function are found in many cancers. CCN3 acts through the core stem cell signalling pathways including Notch and Bone Morphogenic Protein, connecting CCN3 with the modulation of self-renewal and maturation of a number of cell lineages including hematopoietic, osteogenic and chondrogenic. CCN3 may affect the extracellular environment of the niche for hematopoietic stem cells. CCN3 has emerged as a key player in stem cell regulation, hematopoiesis and a crucial component within the bone marrow microenvironment.

References
  • Manara MC, et al. (2002) The expression of ccn3(nov) gene in musculoskeletal tumors. Am J Pathol. 160(3): 849-59.
  • Lin CG, et al. (2003) CCN3 (NOV) is a novel angiogenic regulator of the CCN protein family. J Biol Chem. 278(26): 24200-8.
  • Vallacchi V, et al. (2009) CCN3/nephroblastoma overexpressed matricellular protein regulates integrin expression, adhesion, and dissemination in melanoma. Cancer Res. 68(3): 715-23.
  • Sin WC, et al. (2009) Matricellular protein CCN3 (NOV) regulates actin cytoskeleton reorganization. J Biol Chem. 284(43): 29935-44.
  • McCallum L, et al. (2009) CCN3--a key regulator of the hematopoietic compartment. Blood Rev. 23(2): 79-85.
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    Catalog:SEK10264-5
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    • Human CCN3 / NOV / IGFBP9 ELISA standard curve
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