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 | NoShift II Products • NoShift • Back to Main NoShift Page | | | | The NoShift™ II Transcription Factor Assays enable rapid analysis of transcription factor activity in a convenient 96-well format. This sensitive, non-radioactive assay can be completed within four hours and is a superior alternative to standard electromobility shift assays (EMSA). |
Biotin-labeled DNA probe consisting of a double-stranded consensus transcription factor binding site and a single-stranded capture region is incubated with a nuclear extract. Activated transcription factor binds specifically to the double-stranded consensus sequence. With the addition of a double-stranded DNA specific nuclease, the probe bound to a transcription factor is protected from digestion whereas unbound probe is degraded.
The reactions are then transferred to a 96-well plate, coated with the complementary DNA strand to the capture region of the probe, and the intact probe/transcription factor is captured on the plate. After washing to remove unbound biotin and digested probe, streptavidin-alkaline phosphatase is added and allowed to bind to the biotinylated probe. Another wash step is followed by addition of a chemiluminescent AP substrate. After signal development, detection is performed on any standard microplate luminometer. The output signal is proportional to the amount of probe bound to the transcription factor.
| |  | | The NoShift II assays complement and extend the existing Novagen range of antibody-based NoShift transcription factor analysis tools, and ideal for rapid screening for families of transcription factors based on the specificity of their binding sites. Each assay is internally calibrated, using a control reaction to detect percent digestion of unbound probe, and exhibits excellent reproducibility. | | |
NoShift II
EGR Transcription
Factor Assay | Early growth response gene (EGR1-4) proteins are induced rapidly and transiently upon stimulation. The proteins bind the GCGGGGGCG motif, known as the EGR consensus box. Variously, and in a cell-specific manner, members of the EGR family can be induced during differentiation and by growth factors, LPS, and phorbol esters. | | Cat. No. 71678 | NoShift II
NF-1 Transcription
Factor Assay | The nuclear factor-1 (NF-1) family of transcription factors are encoded by the genes NF-1A, NF-1B, NF-1C, and NF-1X. NF-1 binding controls the expression of many target genes, including insulin, TGF-b, nucleotide, steroid hormone synthetic genes, TNFa, and FSH. After differential splicing of NF-1 transcripts, the resultant protein variants homo- and heterodimerize, allowing for multiple potential transcriptional activation and repression activities. | | Cat. No. 71679 | NoShift II
NF-kB Transcription
Factor Assay | Nuclear factor kB (NF-kB) plays an important role in inflammation, autoimmune response, cell proliferation, and apoptosis by regulating the expression of genes involved in these processes. NF-kB is a heterodimer composed of a combination of two subunits from the p50, p65 (RelA), p52, c-rel, or relB. Several human lymphoid cancer cells are reported to have mutations or amplifications of genes encoding NF-kB subunits. | | Cat. No. 71680 | NoShift II
PPAR Transcription
Factor Assay | Peroxisome proliferator-activated receptors [PPAR a, d (b), and g] are members of the nuclear hormone receptor family of transcription factors that mediate a variety of cellular processes, including glucose and lipid metabolism, inflammatory responses, and regulation of apoptotic cell death. PPARs act by binding to specific peroxisome proliferator-response elements (PPREs) on target genes. | | Cat. No. 71681 |
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