The Non-Radioactive FoxO1 Transcription Factor Assay kit is provided in a 96-well format. During the assay, the Capture Probe, a double stranded biotinylated oligonucleotide containing the consensus sequence for FoxO1 binding (5'-TTGTTTAC-3'), is mixed with whole cell or nuclear extract in the Transcription Factor Assay Buffer provided directly in the Streptavidin coated plate. When incubated together, FoxO1 contained in the nuclear extract binds to its consensus sequence. The biotinylated double stranded oligonucleotide bound to FoxO1 protein is immobilized and any inactive, unbound material is washed away. The bound FoxO1 transcription factor is detected with a specific rabbit anti-FoxO1, primary antibody. An HRP-conjugated secondary antibody is then used for detection and provides sensitive colorimetric detection that can be read in a spectrophotometric plate reader. Included in the kit are positive control whole cell extract, a non-specific double stranded oligonucleotide, and a specific double stranded competitor oligonucleotide.
Applications:
Suitable for use in Western Blot, Immunohistochemistry, and Immunocytochemistry. Other applications not tested.
Recommended Dilution:
Western Blot: 1:500-1000
Immunohistochemistry (paraffin): 1:250-500
Immunocytochemistry: 1:100-250
Optimal dilutions to be determined by the researcher.
Positive Control:
NIH/3T3 cell lysate
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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