Reasons for performing Electrophoretic Mobility Shift Assay (EMSA)
EMSA is a well-established technique for separating macromolecules under native conditions based on a combination of shape, size, charge and interactive molecules. The use of EMSA can provide both general and specific information concerning the interaction between two macromolecules such as DNA/proteins or RNA/proteins (see principle).
Regulation and control of cell transcription factor involved in gene, has a relationship with the organism's illness and death. Normal cells have a normal transcription factor activity, disease is abnormal. Almost every kind of cancer cells have abnormal transcription factor binding. For example, children with lymphoblastic leukemia mostly with aberrant activation of NFkB. Head and neck tumor abnormal activation of STAT3 is very common.
In recent years, the fiery stem cells induced and stem cell technology are related to the role of transcription factors in somatic cells to induced: stem cells into several genes and gene products are all transcription factor.
EMSA is a combination of electrophoresis technology, to be able to identify transcription factor binding. In contrast, other any combination of electrophoretic method for the determination of separation technology, such as gene (DNA) chip, protein chip, micro plate method for determination of the activation of transcription factor binding, transcription factor ChiP were measured, all need to confirm with EMSA. As shown on the right, in the determination of EMSA appeared in 6-7 band. In EMSA test, first and third STAT5 electrophoresis is negative, but in the biological chip dot matrix or microplate assay, there will be a positive signal.
EMSA is the gold standard to confirm the nucleic acid binding protein activity, in a time predictable, EMSA is on the activity of TF was performed to confirm the irreplaceable technology.
Super Shift EMSA is the combination of antibody identification of multi polymer composition. EMSA binding assay to confirm the report gene involved in gene regulation and control.
Determination of activity of transcription factor EMSA is an important part of cell signal transduction pathways, any signal transduction and activation of transcription factors are connected and start the transcription process. Phosphorylation of signal pathway molecules in the sometimes is not nuclear transcription factor EMSA can be detected by immunofluorescence, later proved that nuclear translocation of nuclear NFkB in No.
With the improvement of determination technology, non radioactive EMSA than radioactive isotope labeled (to the results of 4-5 days) faster results than most, and analysis of signal transduction molecules faster results. The use of Viagene's products and technologies (see related technologies and products), EMSA assay can be completed in as little as 2-2.5 hours (view), the sensitivity is more than using radioisotope labeled.