The electrophoretic mobility shift assay emsa, or gel shift assay is a simple and rapid method to detect protein complexes with nucleic acids. If the protein is capable of binding to the fragment, the lane with protein present will contain another band that represents the larger, less mobile complex of nucleic. Protein interaction 2 principle and protocol of emsa. Shop online for a wide selection of thermo scientific lightshift chemiluminescent emsa kit identify and characterize proteindna binding interactions by emsa.
A protocol for electrophoretic mobility shift assay emsa. A general emsa gelshift protocol ran chen department of genetics, stanford university, stanford, usa for correspondence. Electrophoretic mobility shift assay emsa protocol jove. Here, we describe the emsa protocol for analyzing the irebinding activity of irp1 and irp2, which can be generalized to assess the activity of other rnabinding proteins as well.
Dnabinding proteins transcription factors fluorescenceemsa 2demsa gel shift. Cover top of gel with saran wrap and dry at 80 c in. The curriculum, emergency medical technician basic. We relied on the emsa protocol presented here to determine. Electrophoretic mobility shift assay emsa for detecting. Electrophoretic mobility shift assay emsa for the study of rna. The electrophoretic mobility shift assay emsa, or gel shift assay is a simple and rapid method to detect. Procedure to add items to local optional scope of practice. In this protocol, we identify the most important factors that determine the stabilities and electrophoretic mobilities of complexes under assay.
Find, read and cite all the research you need on researchgate. The electrophoretic mobility shiftassay emsa has been used extensively for studying dnaprotein interactions. Multicounty ems agencies using state general fund eff. Fluorescencebased electrophoretic mobility shift assay in the. Solutions of rna and protein are mixed in different ratios, and the binding. National standard curriculum, is the cornerstone of ems prehospital training. Gel shift assay system instructions for use of products e3050 and e3300. The electrophoretic mobility shift assay emsa, one of the most sensitive methods for studying the dnabinding properties of a protein, can be used to deduce.
In addition, the new curriculum parallels the recommendations of the national ems education and practice blueprint. Electrophoretic mobility shift assay emsa for the study. The program can be downloaded from the nih website for free. A schematic representation of a conventional emsa protocol. All software windows mac palm os linux windows 7 windows 8 windows mobile windows phone ios android windows ce windows server pocket pc blackberry tablets os2 handheld symbian openvms unix. The colored boxes in each protocol tree are linked to the respective pages with further information on the specific procedure, medication, or protocol page. The interaction of transcriptional or cotranscriptional factors with dna is crucial for changes of neuronal gene expression during normal brain development as. If viewing this document as portable document file. Protein interaction 2 principle and protocol of emsa creative.
A crude protein lysate containing an rnabinding protein, or a purified preparation of this protein, is incubated with an excess of 32 plabeled rna probe, allowing for complex formation. Emsa originally used widely in the study of sequencespecific dnabinding proteins such. Chapter 19 electrophoretic mobility shift assay for characterizing. Southwest ohio protocol hamilton county oh fire chief. Here, we describe the emsa protocol for analyzing the irebinding activity of irp1 and irp2, which can be generalized.
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