STAT5A (Phospho-Ser780)Transcription Factor Activity Assay

With a 4.5 hour bench workflow, this Assay Genie assay provides all critical reagents in a 12 x 8-well microstrip plate. Each well requires a minimum of 100 µl of sample or diluted sample, with the protocol recommending 5 µg or more of cell lysate per well for reliable quantitation. A Nuclear Lysate Positive Control is included lyophilized and is reconstituted with 110 µl of 1x Binding Buffer prior to use; suggested dilutions are 1:10, 1:20 and 1:40 in 1x Binding Buffer. Samples containing visible precipitate should be centrifuged for 10 minutes at ≥10,000 x g before loading. Color development uses 50 µl of Ready-to-Use Substrate per well for 10 to 30 minutes, followed by 100 µl of Stop Solution, with absorbance measured on a standard microplate reader; readings should be captured within 30 minutes of adding Stop Solution. Component shelf life is 6 months: store the 10x Wash Buffer, Primary Antibody Diluent, Ready-to-Use Substrate and Stop Solution at 4 degrees C, while the 2x Binding Buffer, Nuclear Wash Buffer, Cytoplasmic Extraction Buffer and Nuclear Extraction Buffer must be frozen at -20 degrees C. Intended for research use only.