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Background:
This kit is designed for the detection of cyclin A using the electrophoretic mobility shift assay (EMSA). Applications include screening for the presence of cyclin A in various protein sources, evaluating the functional binding of cyclin A recombinant proteins, analysis of different DNA-binding consensus sites and characterization of promoter elements.Contents:
The following are sufficient for 17 supershift reactions:2 x 40 µl Raji nuclear extract
35 µl cyclin A mouse mAb
40 µl E2F Oligo WT
12 µl E2F Oligo probe WT
40 µl E2F mutant Oligo
240 µl Dilution Buffer A
240 µl Binding Buffer B-4
240 µl Binding Buffer C-1
360 µl Stabilizing Solution D
5 MicroSpin™ G-25 Columns
Application Notes:
When performing EMSA experiments individual optimization of extract and antigen quantities may be required in order to detect the protein of interest. For optimal results, we recommend the use of 5 µg of nuclear extract, 2 µl of antibody and 1X Tris-glycine electrophoresis buffer.Storage:
Upon receipt, the nuclear extract(s) must be stored at -80°C and the Microspin&trade columns should be stored at room temperature. All other components may be stored at either -20°C or -80°C. We recommend aliquoting the extracts into single-use fractions and then storing them at -80°C. This avoids repeated freeze/thaw cycles.Guarantee:
This product is guaranteed for 6 months from date of receipt.This product is for research use only and is not for use in diagnostic procedures.
