NRL DNA-Binding ELISA Kit (OKAG00490)

Catalog No: OKAG00490

Size:96 Wells

NRL DNA-Binding ELISA detects active NRL in HepG2 and HEK293 Nuclear Extract. The HepG2 and HEK293 cells were grown 3 days in DMEM with 10% FBS and harvested for nuclear extract.

Product Info

• Predicted Species Reactivity: Human, Mouse
• Product Format: 12 x 8-Well Microstrips
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 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: 162080
  UniGene: Hs.652297
  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 NRL DNA-Binding ELISA detects endogenous levels of NRL.
• Assay Info: Assay Type: Qualitative DNA Binding ELISA

Target Info

• Gene Symbol: NRL
• Gene Full Name: neural retina leucine zipper
• Alias Symbols: RP27, D14S46E, NRL-MAF
• NCBI Gene Id: 4901
• Protein Name: neural retina-specific leucine zipper protein
• Description of Target: This gene encodes a basic motif-leucine zipper transcription factor of the Maf subfamily. The encoded protein is conserved among vertebrates and is a critical intrinsic regulator of photoceptor development and function. Mutations in this gene have been associated with retinitis pigmentosa and retinal degenerative diseases.
• Uniprot ID: P54845