Size:96 Wells
$539.00
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SKU
OKAG00400
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GR DNA-Binding ELISA Kit (OKAG00400)

Catalog#: OKAG00400
Domestic: within 1 week delivery | International: 1 week
More Information
Predicted Species Reactivity Human, Mouse, Rat
Product Format 12 x 8-Well Microstrips
Application ELISA
Kit Detection Method Colorimetric, OD450 nm
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, or2N 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.
Kit Component
ComponentsAmount
dsDNA Oligonucleotide Coated Microplate96 Wells (12 x 8 Strips)Microstrips
100X Primary Antibody100 uL
100X Primary Phospho-Antibody100 uL
HRP-Conjugated Anti-Rabbit IgG Antibody2 x 6 mL
Nuclear Lysate Positive ControlLyophilized
Wild-Type Consensus dsDNA Oligonucleotide20 uL
Mutant Consensus dsDNA Oligonucleotide20 uL
10X Wash Buffer50 mL
2X Binding Buffer12 mL
Primary Antibody Diluent12 mL
100X Protease and Phosphatase Inhibitors100 uL
Nuclear Wash Buffer12 mL
Cytoplasmic Extraction Buffer6 mL
Nuclear Extraction Buffer6 mL
Ready-to-Use Substrate12 mL
Stop Solution12 mL
:: OMIM: 138040
UniGene: Hs.122926
:: 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.
Datasheets/Manuals Click here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Specificity The GR DNA-Binding ELISA Kit detects endogenous levels of total GR protein.
Assay Info Assay Type: Qualitative DNA Binding ELISA
Gene Symbol NR3C1
Official Gene Full Name nuclear receptor subfamily 3 group C member 1
Alias Symbols GCCR, GCR, Glucocorticoid receptor, GR, GRL, Nuclear receptor subfamily 3 group C member 1
NCBI Gene Id 2908
Protein Name glucocorticoid receptor
Description of Target This gene encodes glucocorticoid receptor, which can function both as a transcription factor that binds to glucocorticoid response elements in the promoters of glucocorticoid responsive genes to activate their transcription, and as a regulator of other transcription factors. This receptor is typically found in the cytoplasm, but upon ligand binding, is transported into the nucleus. It is involved in inflammatory responses, cellular proliferation, and differentiation in target tissues. Mutations in this gene are associated with generalized glucocorticoid resistance. Alternative splicing of this gene results in transcript variants encoding either the same or different isoforms. Additional isoforms resulting from the use of alternate in-frame translation initiation sites have also been described, and shown to be functional, displaying diverse cytoplasm-to-nucleus trafficking patterns and distinct transcriptional activities (PMID:15866175).
Swissprot Id P04150
Tissue Tool Find tissues and cell lines supported by DNA array analysis to express NR3C1.
RNA Seq Find tissues and cell lines supported by RNA-seq analysis to express NR3C1.
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