STAT3 DNA-Binding ELISA Kit (OKAG00467)

Catalog No: OKAG00467

Size:12x8WELLSTRIPS

STAT3 DNA-Binding ELISA detects active STAT3 in Hela and HepG2 Nuclear Extract. The HepG2 and Hela cells were grown 3 days in DMEM with 10% FBS and harvested for nuclear extract. The Hela cells were stimulated by PMA (200nM) before harvest.

Product Info

• Predicted Species Reactivity: Human, Mouse, Rat
• Product Format: 12 x 8-Well Microstrips
• Application: Enzyme-linked immunosorbent assay
• ELISA Kit Detection Method: Colorimetric 450 nm
• ELISA Kit Principle: The Aviva DNA-Binding ELISA Kit contains components necessary for detection of active transcription factors in eukaryotic nuclear or cell lysates. This particular immunoassay utilizes the qualitative technique of an indirect ELISA. Streptavidin is bound to the immunoassay plate and specific biotinylated double-stranded (dsDNA) oligonucleotides are then added to bind to the streptavidin via a high affinity biotin-streptavidin interaction. After subsequent blocking of extraneous binding sites in each well, the sample containing the target of interest can be added. Primary antibody is added to bind activated transcription factors bound to the dsDNA oligonucleotide, which has been immobilized via the plate-coated streptavidin. A HRP-conjugated secondary antibody specific for rabbit IgGs is added, which allows for specific binding to the Primary Antibody, and consequently colorimetric detection upon addition of the TMB substrate.
  For color development, TMB (3, 3’, 5, 5’-Tetramethylbenzidine) is added to each well. After addition of the substrate, a peroxidase catalyzed reaction will produce a blue TMB Diimine product that is proportional to the target concentration in the sample. Color development is quenched by addition of Stop Solution, or 2 N Sulfuric Acid, which turns the solution yellow. The absorbance can then be read by a spectrophotometer at 450 nm and subsequently allowing for determination of the target concentration in the sample.
  
• ELISA Kit Component:
  • dsDNA Oligonucleotide Coated Microplate: 96 Wells (12 x 8 Strips)Microstrips
  • 100X Primary Antibody: 100 uL
  • 100X Primary Phospho-Antibody: 100 uL
  • HRP-Conjugated Anti-Rabbit IgG Antibody: 2 x 6 mL
  • Nuclear Lysate Positive Control: Lyophilized
  • Wild-Type Consensus dsDNA Oligonucleotide: 20 uL
  • Mutant Consensus dsDNA Oligonucleotide: 20 uL
  • 10X Wash Buffer: 50 mL
  • 2X Binding Buffer: 12 mL
  • Primary Antibody Diluent: 12 mL
  • 100X Protease and Phosphatase Inhibitors: 100 uL
  • Nuclear Wash Buffer: 12 mL
  • Cytoplasmic Extraction Buffer: 6 mL
  • Nuclear Extraction Buffer: 6 mL
  • Ready-to-Use Substrate: 12 mL
  • Stop Solution: 12 mL
  
  OMIM: 102582/147060
  UniGene: Hs.463059
  Note: The product is for research use only, not for use in diagnostic or therapeutic procedures.
• Reconstitution and Storage: Store as indicated in product manual.
• Specificity: The STAT3 DNA-Binding ELISA detects endogenous levels of total STAT3 protein.
• Assay Info: Assay Type: Qualitative DNA Binding ELISA

Target Info

• Gene Symbol: STAT3
• Gene Full Name: signal transducer and activator of transcription 3
• Alias Symbols: APRF, HIES, ADMIO, ADMIO1
• NCBI Gene Id: 6774
• Protein Name: signal transducer and activator of transcription 3
• Description of Target: The protein encoded by this gene is a member of the STAT protein family. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor associated kinases, and then form homo- or heterodimers that translocate to the cell nucleus where they act as transcription activators. This protein is activated through phosphorylation in response to various cytokines and growth factors including IFNs, EGF, IL5, IL6, HGF, LIF and BMP2. This protein mediates the expression of a variety of genes in response to cell stimuli, and thus plays a key role in many cellular processes such as cell growth and apoptosis. The small GTPase Rac1 has been shown to bind and regulate the activity of this protein. PIAS3 protein is a specific inhibitor of this protein. Mutations in this gene are associated with infantile-onset multisystem autoimmune disease and hyper-immunoglobulin E syndrome. Alternative splicing results in multiple transcript variants encoding distinct isoforms.
• Uniprot ID: P40763