K-025
NFkB Oligonucleotide
Cannot supply to this region.
- SKU:
- K-025
- Additional Names:
- NFkB NFkappaB
- Application:
- ELISA, WB
- Concentration:
- 20 ng/ul
- Physical State:
- Liquid
- Storage Conditions:
- -20[o]C aliquoted. Aliquot. Avoid freeze/thaw cycles., 2-8[o]C diluted. Aliquot. Avoid freeze/thaw cycles.
- Supplier:
- Rockland Inc
- Buffer:
- 0.01 M Tris Cl, 0.15 M Sodium Chloride, 0.001 M EDTA
- Formulation:
- 0.01 M Tris Cl, 0.15 M Sodium Chloride, 0.001 M EDTA, pH 7.4
- Extra Details:
- NFkB oligonucleotide consensus sequence for binding of NFkB /c-Rel homodomeric and heterodimeric complexes. Supplied as 500 ng of double stranded DNA for gel shift and super shift assays. In general, NFkB gel shift assays are assembled in 20ul reactions containing 0.28pmoles NFkB oligo in 10mM Tris (pH 7.6), 50 mM NaCl, 0.5 mM EDTA, 1.0 mM DTT, 10% glycerol. Some procedures specify the addition of 0.05% NP-40. When using purified protein, 250-300 ng should be sufficient to produce a gel shifted complex, while 10ug HeLa nuclear extract is utilized. The gel shift reactions are then incubated at room temperature for 30 minutes. The complexes are resolved on a Tris-Glycine acrylamide gels. Loading dye containing bromophenol blue and xylene cyanol should be added to the negative control reaction only, as these dyes can increase the dissociation of the NFkB complexes. When using HeLa nuclear extract as the source of binding proteins, two sequence-specific gel-shifted complexes are expected, consisting of p50/p50 homodimers and p50/p65 heterodimers. For cells expressing p52, p50, and p65, as many as four sequence-specific gel-shifted complexes could be observed (p52/p52, p50/p50, p52/p65, p50/p65), and if high levels of p65 are present, the p65/p65 homodimer may also be weakly detected. The following reagents have been observed to enhance NF-kB binding in vitro: millimolar amounts of GTP and ATP, spermine, spermidine, barium or calcium ions, and uM amounts of Co+3(NH3)6.
- Shipping Conditions:
- Dry Ice