Efficient Mammalian Cell Expression and Single-step Purification of Extracellular Glycoproteins for Crystallization

13.7K Views

•

07:08 min

•

December 23rd, 2015

DOI :

10.3791/53445-v

December 23rd, 2015

•

Daniel L. Kober*¹^,²^,³, Zeynep Yurtsever*²^,³^,⁴^,⁵, Thomas J. Brett²^,³^,⁵^,⁶^,⁷

¹Molecular Microbiology and Microbial Pathogenesis Program, Washington University School of Medicine, ²Department of Internal Medicine, Washington University School of Medicine, ³Drug Discovery Program in Pulmonary and Critical Care Medicine, Washington University School of Medicine, ⁴Biochemistry Program, Washington University School of Medicine, ⁵Center for the Investigation of Membrane Excitability Diseases, Washington University School of Medicine, ⁶Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, ⁷Department of Cell Biology and Physiology, Washington University School of Medicine

Summary

Explore More Videos

Keywords Mammalian Cell Expression

Extracellular Glycoproteins

Crystallization

Transient Transfection

Nickel Affinity Chromatography

Protein Production

Protein Yield

Glycan Maturation

X ray Crystallography

Chapters in this video

0:05

Title

1:02

Large-scale Culture and Transient Transfection of 293F Cells

2:40

Purification

4:58

Results: Expression and Purification of Secreted Mammalian Proteins

6:29

Conclusion

Related Videos

08:15

The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation

25.7K Views

07:47

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

53.0K Views

10:17

Pulse-chase Analysis of N-linked Sugar Chains from Glycoproteins in Mammalian Cells

15.2K Views

15:31

MISSION esiRNA for RNAi Screening in Mammalian Cells

16.2K Views

14:40

Expression Analysis of Mammalian Linker-histone Subtypes

13.7K Views

07:50

Efficient Agroinfiltration of Plants for High-level Transient Expression of Recombinant Proteins

48.5K Views

13:52

Efficient Production and Purification of Recombinant Murine Kindlin-3 from Insect Cells for Biophysical Studies

23.3K Views

14:53

Analysis of Global RNA Synthesis at the Single Cell Level following Hypoxia

12.3K Views

10:43

siRNA Screening to Identify Ubiquitin and Ubiquitin-like System Regulators of Biological Pathways in Cultured Mammalian Cells

11.3K Views

09:43

Measurement of Heme Synthesis Levels in Mammalian Cells

12.0K Views