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Catalog No: OKAG00423
Size:12x8WELLSTRIPS
Price: $600.00
SKU
OKAG00423
Availability: Domestic: within 1-2 week delivery | International: 1-2 weeks
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Datasheets/ManualsClick here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Product Info
Predicted Species ReactivityHuman, Mouse, Rat
Product Format12 x 8-Well Microstrips
ApplicationELISA
ELISA Kit Detection MethodColorimetric 450 nm
ELISA Kit PrincipleThe 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
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: 164014
UniGene: Hs.502875
::Note: The product is for research use only, not for use in diagnostic or therapeutic procedures.
Reconstitution and StorageStore as indicated in product manual.
SpecificityThe NF-kappaB p65 DNA-Binding ELISA Kit detects endogenous levels of total NF-kappaB p65 protein.
Assay InfoAssay Type: Qualitative DNA Binding ELISA
Gene SymbolRELA
Gene Full NameRELA proto-oncogene, NF-kB subunit
Alias Symbolsp65, CMCU, NFKB3
NCBI Gene Id5970
Protein Nametranscription factor p65
Description of TargetNF-kappa-B is a ubiquitous transcription factor involved in several biological processes. It is held in the cytoplasm in an inactive state by specific inhibitors. Upon degradation of the inhibitor, NF-kappa-B moves to the nucleus and activates transcription of specific genes. NF-kappa-B is composed of NFKB1 or NFKB2 bound to either REL, RELA, or RELB. The most abundant form of NF-kappa-B is NFKB1 complexed with the product of this gene, RELA. Four transcript variants encoding different isoforms have been found for this gene.
Uniprot IDQ04206
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