Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins

24.0K Views

•

10:21 min

•

June 20th, 2019

DOI :

10.3791/59729-v

June 20th, 2019

•

Alexander K. H. Weiss¹^,², Max Holzknecht¹^,², Elia Cappuccio¹^,², Ilaria Dorigatti¹, Karin Kreidl¹, Andreas Naschberger³, Bernhard Rupp³, Hubert Gstach⁴, Pidder Jansen-Dürr¹^,²

¹Research Institute for Biomedical Aging Research, University of Innsbruck Austria, ²Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck Austria, ³Division of Genetic Epidemiology, Medical University of Innsbruck Austria, ⁴Faculty of Chemistry, Department of Organic Chemistry, University of Vienna Austria

Summary

Explore More Videos

Fumarylacetoacetate Hydrolase

Affinity Chromatography

IPTG Inducible Vector

Protein Quantity Amplification

Chapters in this video

0:04

Title

1:12

Expression of FAHD Proteins in Competent E. coli

3:44

Lysis of Bacterial Pellets and Filtration of Debris

4:33

Purification of His-tagged FAHD Proteins using Ni-NTA Affinity Chromatography

5:59

Purification of FAHD Proteins via Ion Exchange Chromatography

6:35

Basic FAHD Activity Assays with Substrates Oxaloacetate and Acetylpyruvate

7:53

Results: Expression and Purification of Fumarylacetoacetate Hydrolase Domain-containing Proteins

9:19

Conclusion

Related Videos

12:21

Thermostabilization, Expression, Purification, and Crystallization of the Human Serotonin Transporter Bound to S-citalopram

15.0K Views

11:14

Combining Wet and Dry Lab Techniques to Guide the Crystallization of Large Coiled-coil Containing Proteins

7.9K Views

11:10

Expression, Purification, and Antimicrobial Activity of S100A12

8.3K Views

12:23

Recombinant Protein Expression, Crystallization, and Biophysical Studies of a Bacillus-conserved Nucleotide Pyrophosphorylase, BcMazG

7.4K Views

08:12

Expression and Purification of Mammalian Bestrophin Ion Channels

8.3K Views

09:14

Modifying Baculovirus Expression Vectors to Produce Secreted Plant Proteins in Insect Cells

11.4K Views

08:55

Expression, Solubilization, and Purification of Eukaryotic Borate Transporters

9.5K Views

10:14

Expression and Purification of Nuclease-Free Oxygen Scavenger Protocatechuate 3,4-Dioxygenase

6.3K Views

05:22

Transverse Sectioning of Mature Rice (Oryza sativa L.) Kernels for Scanning Electron Microscopy Imaging Using Pipette Tips as Immobilization Support

3.5K Views

09:31

PCR Mutagenesis, Cloning, Expression, Fast Protein Purification Protocols and Crystallization of the Wild Type and Mutant Forms of Tryptophan Synthase

4.2K Views