Androgen Receptor DNA-Binding ELISA Kit (OKAG00363)

Catalog No: OKAG00363

Size:12x8WELLSTRIPS

Androgen Receptor DNA-Binding ELISA detects active Androgen Receptor in MCF7 Nuclear Extract. MCF7 cells were grown 3 days in DMEM with 10% FBS and harvested for nuclear extract.

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 2N Sulfuric Acid, which turns the solution yellow. The absorbance can then be read by a spectrophotometer at 450nm 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: 109200/300068/312300/313200/313700
  UniGene: Hs.496240
  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 Androgen Receptor DNA-Binding ELISA Kit detects endogenous levels of total Androgen Receptor protein.
• Assay Info: Assay Type: Qualitative DNA Binding ELISA

Target Info

• Gene Symbol: AR
• Gene Full Name: androgen receptor
• Alias Symbols: KD, AIS, AR8, TFM, DHTR, SBMA, HYSP1, NR3C4, SMAX1, HUMARA
• NCBI Gene Id: 367
• Protein Name: androgen receptor
• Description of Target: The androgen receptor gene is more than 90 kb long and codes for a protein that has 3 major functional domains: the N-terminal domain, DNA-binding domain, and androgen-binding domain. The protein functions as a steroid-hormone activated transcription factor. Upon binding the hormone ligand, the receptor dissociates from accessory proteins, translocates into the nucleus, dimerizes, and then stimulates transcription of androgen responsive genes. This gene contains 2 polymorphic trinucleotide repeat segments that encode polyglutamine and polyglycine tracts in the N-terminal transactivation domain of its protein. Expansion of the polyglutamine tract causes spinal bulbar muscular atrophy (Kennedy disease). Mutations in this gene are also associated with complete androgen insensitivity (CAIS). Two alternatively spliced variants encoding distinct isoforms have been described.
• Uniprot ID: P10275