Bioinformatics, Protocols, DNA RNA Protein Proteomics
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In a manner which matches the fortuity, if not the consequence, of Archimedes' bath and Newton's apple, the [3.6 million year old] fossil footprints were eventually noticed one evening in September 1976 by the paleontologist Andrew Hill, who fell while avoiding a ball of elephant dung hurled at him by the ecologist David Western. ~John Reader, Missing Links: The Hunt for Earliest Man
EMSA Protocols for DNA-binding protein assessment using Electrophoretic Mobility Shift Assay.
| A non-radioactive electrophoretic mobility shift assay for the detection of heat shock element (HSE) - http://www.fgsc.net/fgn45/45meyer.html A non-radioactive electrophoretic mobility shift assay based on the digoxigenin detection system had been developed and applied to the analysis of the interaction between the heat shock transcription factor and the heat shock element of N. crassa. (Bioch - [Read A non-radioactive electrophoretic mobility shift assay for the detection of heat shock element (HSE)] |
| DNA-Binding Protein Purification with Dynabeads. - http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=2671937 Magnetic DNA affinity purification of yeast transcription factor tau--a new purification principle for the ultrarapid isolation of near homogeneous factor. Gabrielsen et al. 1989 - [Read DNA-Binding Protein Purification with Dynabeads.] |
| DNaseI Footprinting Assay - http://www.fhcrc.org/science/labs/breeden/Methods/DNaseI-footprint.html DNaseI Footprinting Assay. Works well high affinity dna binding proteins. Breeden Lab. - [Read DNaseI Footprinting Assay] |
| Electrophoretic Mobility Shift Assay (EMSA) for the detection of nuclear NF-kB - http://www.celldeath.de/apometh/emsa.html Electrophoretic Mobility Shift Assay (EMSA) for the detection of nuclear NF-kB. CellDeath.de - [Read Electrophoretic Mobility Shift Assay (EMSA) for the detection of nuclear NF-kB] |
| EMSA protocol (Word Doc) - http://www.med.umich.edu/intmed/endocrinology/hammerlab/Protocols/EMSA%20Protocol.doc Short EMSA protcol starting with gel running and supershift assay. Hammer Lab. - [Read EMSA protocol (Word Doc)] |
| EMSA Protocol using 32-p. - http://www.well.ox.ac.uk/flint/EMSA.htm Primer design, Primer annealing, Primer labelling, Probe purification, In vitro hybridization DNA-protein Binding reaction. Jonathan Flint. - [Read EMSA Protocol using 32-p.] |
| EMSA using ds Oligonucleotides - http://axon.med.harvard.edu/~cepko/protocol/mike/E1.html EMSA probe creation using ds Oligonucleotides by annealing two complementary oligos, and Klenow for probe creation. - [Read EMSA using ds Oligonucleotides] |
| Gel Mobility Shift Assay in PDF format - http://www.biochem.northwestern.edu/ibis/morimoto/Protocols/III.%20Proteins/C.%20DNA-Protein%20Interactions/3a.%20GMSA.pdf This protocol works well for HSF DNA-binding activity experiments. - [Read Gel Mobility Shift Assay in PDF format] |
| General EMSA Protocol - http://faculty.virginia.edu/mirmira/Protocols/emsa.html There are multiple variations to this protocol, but they find that this one works well in all cases tested. Mirmira Lab. - [Read General EMSA Protocol] |
| Introduction to the EMSA (Gel Shift) Technique - http://www.piercenet.com/Proteomics/browse.cfm?fldID=7EDAC33E-3981-4E58-8A57-057A8280C68F A non-radioactive electrophoretic mobility which works quite well using biotin and streptavidin detection. Non-specific and specific competititor, oligo labeling, Binding Reaction. Pierce - [Read Introduction to the EMSA (Gel Shift) Technique] |
| Nuclear Extract Preparation for EMSA (PDF) - http://research.dfci.harvard.edu/shivdasani/protocols/Nuclear_extract_prep.pdf Nuclear Extract Preparation for EMSA after Wadman et al., EMBO J. 1997. Buffer recipies included. - [Read Nuclear Extract Preparation for EMSA (PDF)] |
| Protocol for Non-Radioactive EMSAs Using DIG - http://www.med.umich.edu/intmed/endocrinology/hammerlab/Protocols/Protocol%20for%20Non-radioactive%20EMSA.doc Very Nice and Detailed Protocol for Non-Radioactive EMSAs Using DIGoxin. Hammer Lab. - [Read Protocol for Non-Radioactive EMSAs Using DIG] |
| Radiolabeling of Probes for Electrophoretic Mobility Shift Assay - http://faculty.virginia.edu/mirmira/resources_files/Protocols/Radiolabeling.htm Creation of Probe for EMSA using Oligonucleotide annealing, p-32, and T4 Polynucleotide Kinase. - [Read Radiolabeling of Probes for Electrophoretic Mobility Shift Assay] |
EMSA Electrophoretic Mobility Gel Shift Reaction ProtocolEMSA Electrophoretic Mobility Gel Shift Reaction Protocol. |
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