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CYT420 | ChemiKine™ Pigment Epithelium Derived Factor, Sandwich ELISA

CYT420
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      Overview

      Replacement Information

      Key Specifications Table

      Analytes AvailableSpecies ReactivityKey ApplicationsDetection Methods
      PEDF H ELISA Colorimetric
      Description
      Catalogue NumberCYT420
      Brand Family Chemicon®
      Trade Name
      • ChemiKine
      • Chemicon
      DescriptionChemiKine™ Pigment Epithelium Derived Factor, Sandwich ELISA
      OverviewIntroduction

      Pigment Epithelium-Derived Factor (PEDF) was first identified as a factor secreted by primary retinal pigment epithelial (RPE) cells that promotes the survival of neurons in vitro and in vivo. Molecular characterization of PEDF shows it to be a 50 kDa member of the serpin family of protease inhibitors1. A second mode of action of PEDF was identified with the demonstration that PEDF induces apoptosis of microvascular endothelial cells, a property that maintains the avascular nature of the retina2. As a number of ophthalmologic disorders are characterized by dysregulation of the retinal innervation and microvasculature, PEDF serves as an important regulator of human eye disease3. In addition, PEDF appears to have multifunctional antitumor activity in a neuroblastoma model, as Schwann cell-derived PEDF induces a differentiated, less malignant phenotype in neuroblastoma cells, promotes further growth and survival of Schwann cells, and inhibits angiogenesis4.

      ChemiKine™ Pigment Epithelium-Derived Factor (PEDF) kit is a sandwich stripwell-type enzyme immunoassay (EIA), which measures PEDF. The kit will measure PEDF of vitreous fluid of human origin.

      Test Principle

      With the ChemiKine™ PEDF assay system, a mouse monoclonal antibody generated against human PEDF is coated onto a microplate and is used to capture PEDF from a sample. A second, biotinylated PEDF-specific mouse monoclonal antibody detects the captured PEDF. Peroxidase-labeled streptavidin detects the biotinylated mouse monoclonal antibody. After addition of the substrate and stop solution the amount of PEDF is determined. The standard curve demonstrates a direct relationship between Optical Density (OD) and PEDF concentration: i.e., the higher the OD the higher the PEDF concentration in the sample.
      Alternate Names
      • PEDF
      Materials Required but Not Delivered1. Multi-channel or repeating pipettes

      2. 37ºC incubator

      3. Plate sealers

      4. Pipettors & tips capable of accurately measuring 10-1000 mL

      5. Graduated serological pipettes

      6. 96-well microplate reader with 450 nm filter

      7. Graph paper for manual plotting of data

      8. Polypropylene test tubes for standard and sample dilutions

      9. Mechanical vortex

      10. One 1 or 2 liter container

      11. Urea
      References
      Product Information
      Components
      • PEDF ELISA Plate: (Part No. 61597) One 96-Well Immunoplate Pre-coated with Mouse anti-Human PEDF Monoclonal Antibody, sealed in a foil pouch.
      • Wash Buffer Concentrate: (Part No. 60245) One 100 mL (10X) bottle of Concentrate.
      • Assay Diluent: (Part No. 60240) One 60 mL bottle (Ready to Use).
      • PEDF Standard (Human): (Part No. 61600) One vial containing 250 ng recombinant full length human PEDF (Lyophilized).
      • Biotinylated Mouse anti-Human PEDF Monoclonal Antibody: (Part No. 61598) One 50 μL vial.
      • Streptavidin, HRP conjugated: (Part No. 61599) One 50 μL vial.
      • TMB/E Solution: (Part No. 60096) One 10 mL bottle of a Ready to Use solution of 3,3',5,5'-tetramethylbenzidine in a proprietary buffer with enhancer.
      • Stop Solution (Part No. 60193): One 12 mL bottle of an HCl solution (Ready to Use).
      Detection methodColorimetric
      Applications
      ApplicationThis ChemiKine Pigment Epithelium Derived Factor, Sandwich ELISA is used to measure & quantify PEDF levels in Neuroscience research.
      Key Applications
      • ELISA
      Application NotesChemiKine™ PEDF kit is designed to measure the amount of PEDF in cell culture supernatants, tissue homogenates and biological fluid (vitreous, serum, plasma, and serum-free) samples of human origin. There are enough reagents included in this kit for one 96-well immunoassay plate. Running duplicate wells for samples and standards is recommended.

      For Research Use Only. Not for use in diagnostic procedures.
      Biological Information
      HostMouse
      Species Reactivity
      • Human
      Antibody TypeMonoclonal Antibody
      Analytes Available
      • PEDF
      Entrez Gene Number
      Gene Symbol
      • SERPINF1
      • Serpin-F1
      • PEDF
      • PIG35
      • EPC-1
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P36955 # Neurotrophic protein; induces extensive neuronal differentiation in retinoblastoma cells. Potent inhibitor of angiogenesis. As it does not undergo the S (stressed) to R (relaxed) conformational transition characteristic of active serpins, it exhibits no serine protease inhibitory activity.
      SIZE: 418 amino acids; 46342 Da
      SUBCELLULAR LOCATION: Secreted. Melanosome. Note=Enriched in stage I melanosomes.
      TISSUE SPECIFICITY: Retinal pigment epithelial cells and blood plasma.
      DEVELOPMENTAL STAGE: Expressed in quiescent cells.
      DOMAIN: SwissProt: P36955 The N-terminal (AA 44-121) exhibits neurite outgrowth- inducing activity. The C-terminal exposed loop (AA 382-418) is essential for serpin activity.
      PTM: The N-terminus is blocked.
      SIMILARITY: Belongs to the serpin family.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsMaintain the unopened kit at 2-8°C until expiration date indicated on the label. After opening the kit maintain each component at 2-8°C until expiration date indicated on the label. After reconstitution, maintain the PEDF Standard at -20ºC for up to 30 days. The standard may be frozen and thawed three times without loss of immunoreactivity. Do not store reconstituted standard at 4ºC, as loss of immunoreactivity will result.

      Precautions

      · Wash Buffer and Assay Diluent contain thimerosal. Thimerosal is highly toxic by inhalation, contact with skin or if swallowed. Thimerosal is a possible mutagen and should be handled accordingly.

      · The instructions provided have been designed to optimize the kit's performance. Deviation from the instructions may result in suboptimal performance of the kit and the failure to produce accurate data.
      Packaging Information
      Material Size1 kit
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      ChemiKine ™ Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit - 2451401 2451401
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit - 2103919 2103919
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit - 2366544 2366544
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit - 2387788 2387788
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit - 2395694 2395694
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit - 2430385 2430385
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit - 2436387 2436387
      ChemiKineTM -2742286 2742286
      ChemiKineTM -2803211 2803211
      ChemiKineTM -2811928 2811928
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit - 2135694 2135694
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit - 2135694A 2135694A
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit - 2212037 2212037
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit - 2290955 2290955
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit -2465210 2465210
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit -2554894A 2554894A
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit- 2549406A 2549406A
      ChemiKineTM Pigment Epithelium-Derived Factor (PEDF) Sandwich ELISA Kit- 2565741A 2565741A
      ChemiKineTM Pigment Epithelium-Derived Factor - 2459594 2459594

      References

      Reference overviewPub Med ID
      Effect of over-expression of pigment epithelium derived factor (PEDF) on developing retinal vasculature in the mouse
      Wong, Wai T, et al
      Mol Vis, 10:837-44 (2004) 2004

      15547489 15547489
      Suppression of corneal neovascularization by PEDF release from human amniotic membranes.
      Shao, Chunkui, et al.
      Invest. Ophthalmol. Vis. Sci., 45: 1758-62 (2004) 2004

      15161837 15161837
      Derivation and comparative assessment of retinal pigment epithelium from human embryonic stem cells using transcriptomics
      Klimanskaya, I. et al.
      Cloning and Stem Cells, 6:217-245 (2004) 2004

      15671670 15671670

      Brochure

      Title
      Immunoassay and Multiplex Kits: Analyte Quarterly Vol. 2 (EMD)
      A closer look at immunoassay platforms. Beyond fit for purpse. Bring your biomarkers to life with EMD Millipore Immunoassay Platform Solutions.
      ELISAs, RIAs & GyroMark HT Assays
      Protein Detection Platform and Services Brochure

      Data Sheet

      Title
      ChemiKine Pigment Epithelium Derived Factor, Sandwich ELISA - Data Sheet
      ChemiKine Pigment Epithelium Derived Factor, Sandwich ELISA - Data Sheet
      Full Service Custom-built Assays

      Posters

      Title
      Multiplex Assays of Fifteen Human Complement Factors

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      Categories

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      Life Science Research > Protein Detection and Quantification > Immunoassays > Enzyme-linked Immunosorbent Assay (ELISA) > Complete ELISA Kits