Temperature Shift Assay
Temperature Shift Assay - Related Videos
06:57
Electrophoretic Mobility Shift Assay (EMSA)
0 Views •
2023
The electrophoretic mobility shift assay (EMSA) is a biochemical procedure used to elucidate binding between proteins and nucleic acids. In this assay a radiolabeled nucleic acid and test protein are mixed. Binding is determined via gel electrophoresis which separates components based on mass, charge, and conformation. This video shows the concepts of EMSA and a general procedure, including gel and protein preparation, binding, electrophoresis, and detection. Applications covered in this video...
03:58
Temperature Gradient Assay: A Method to Test Temperature Preference in Drosophila Larvae
0 Views •
2023
Drosophila fruit flies can distinguish small changes in temperature and therefore select environments with favorable thermal conditions. This video describes a behavioral assay that tests the temperature preference of Drosophila larvae on a linear thermal gradient.
04:31
Electrophoretic Mobility Shift Assay for Detection of Transcription Factor-DNA Complexes
0 Views •
2025
In this video, we demonstrate the electrophoretic mobility shift assay to detect protein-DNA interactions using infrared fluorescent-labeled DNA probes. When the protein binds to the DNA probe, it forms a large complex that migrates slowly on and appears as a shifted band on the gel.
09:12
Sound Waves and Doppler Shift
0 Views •
2023
Source: Arianna Brown, Asantha Cooray, PhD, Department of Physics & Astronomy, School of Physical Sciences, University of California, Irvine, CA Waves are disturbances that propagate through a material medium or empty space. Light waves can travel through a vacuum and some forms of matter, and are transverse in nature, which means that the oscillations are perpendicular to the direction of propagation. However, sound waves are pressure waves that travel through an elastic medium like air,...
11:35
Screening for Functional Non-coding Genetic Variants Using Electrophoretic Mobility Shift Assay (EMSA) and DNA-affinity Precipitation Assay (DAPA)
0 Views •
Cited by 15 •
2016
We present a strategic plan and protocol for identifying non-coding genetic variants affecting transcription factor (TF) DNA binding. A detailed experimental protocol is provided for electrophoretic mobility shift assay (EMSA) and DNA affinity precipitation assay (DAPA) analysis of genotype-dependent TF DNA binding.
FAQs
At the permissive temperature, a temperature-sensitive protein can function normally, establishing a baseline for comparison. Moving the system to the restrictive temperature changes the protein’s folding or stability, which can expose the consequences of altered activity. Comparing function across these conditions helps connect a specific protein state with a cellular or organismal phenotype.
The assay uses temperature as a controlled way to alter the behavior of a temperature-sensitive protein. At the restrictive temperature, changes in folding or stability can disrupt the protein’s normal contribution to a biological process. This relationship allows researchers to associate temperature-dependent molecular changes with outcomes such as altered cell-cycle progression, trafficking, or development.
The timing of the shift provides temporal control over when a temperature-sensitive protein is affected. Observing the resulting change after that point can help determine when the protein is required during a biological process. This is especially informative for processes that unfold in stages, including cell-cycle progression, protein trafficking, and development.
A typical workflow begins with cells or organisms maintained at a permissive temperature, followed by transfer to a restrictive temperature. Researchers then monitor the relevant cellular, molecular, or organismal outcome and may return the system to the permissive condition to assess recovery. The sequence links temperature changes with functional consequences over time.
Temperature shift assays can help identify essential genes and determine when their associated proteins act. They also support studies of cell-cycle progression, protein trafficking, and development by temporarily changing protein function and observing the resulting phenotype. These applications make the method useful for connecting molecular activity to broader biological events.
Returning cells or organisms to the permissive temperature provides an additional test after the restrictive phase. Researchers can examine whether the biological system shows recovery and compare that result with the earlier phenotype. This follow-up adds information about the relationship between the temperature-sensitive protein’s condition and the observed cellular, molecular, or organismal function.