Protein Purification-free Method of Binding Affinity Determination by Microscale Thermophoresis

30.4K Views

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10:22 min

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August 15th, 2013

DOI :

10.3791/50541-v

August 15th, 2013

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Lyuba Khavrutskii¹^,², Joanna Yeh¹, Olga Timofeeva³, Sergey G. Tarasov⁴, Samuel Pritt¹, Karen Stefanisko¹, Nadya Tarasova¹

¹Cancer and Inflammation Program, National Cancer Institute, ²Basic Science Program, SAIC-Frederick, Inc., ³Departments of Oncology and Radiation Medicine, Georgetown University Medical Center, ⁴Structural Biophysics Laboratory, National Cancer Institute

Summary

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Protein Purification

Microscale Thermophoresis

Non denaturing Conditions

Drug Discovery

Protein Interactions

Chapters in this video

0:05

Title

1:43

Preparation of Cell Lysate

3:20

Determination of Optimal Lysate Dilution

4:02

Microscale Thermophoresis Binding Studies

7:05

Microscale Thermophoresis Data Analysis

8:25

Results: Measuring the Affinity of Non-phosphorylated STAT3 Protein Binding to Oligonucleotides

9:40

Conclusion

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