التفاصيل البيبلوغرافية
| العنوان: |
High-Throughput Cellular Thermal Shift Assay Using Acoustic Transfer of Protein Lysates |
| المؤلفون: |
Ashley E. Owens (12046547), Michael J. Iannotti (6334349), Tino W. Sanchez (1936552), Ty Voss (12046550), Abhijeet Kapoor (199911), Matthew D. Hall (316516), Juan J. Marugan (191299), Sam Michael (105053), Noel Southall (187625), Mark J. Henderson (811537) |
| سنة النشر: |
2022 |
| المجموعة: |
Smithsonian Institution: Digital Repository |
| مصطلحات موضوعية: |
Biochemistry, Cell Biology, Genetics, Molecular Biology, Pharmacology, Biotechnology, Cancer, Inorganic Chemistry, Biological Sciences not elsewhere classified, Chemical Sciences not elsewhere classified, preclude screening applications, complex cellular environment, phase protein arrays, throughput cetsa using, protein ’, well microplates, valuable method, thermal stability, limit throughput, lactate dehydrogenase, identify compounds, engage mek1, detection modality, detecting changes, acoustic rppa |
| الوصف: |
Cellular thermal shift assay (CETSA) is a valuable method to confirm target engagement within a complex cellular environment, by detecting changes in a protein’s thermal stability upon ligand binding. The classical CETSA method measures changes in the thermal stability of endogenous proteins using immunoblotting, which is low-throughput and laborious. Reverse-phase protein arrays (RPPAs) have been demonstrated as a detection modality for CETSA; however, the reported procedure requires manual processing steps that limit throughput and preclude screening applications. We developed a high-throughput CETSA using an acoustic RPPA (HT-CETSA-aRPPA) protocol that is compatible with 96- and 384-well microplates from start-to-finish, using low speed centrifugation to remove thermally destabilized proteins. The utility of HT-CETSA-aRPPA for guiding structure–activity relationship studies was demonstrated for inhibitors of lactate dehydrogenase A. Additionally, a collection of kinase inhibitors was screened to identify compounds that engage MEK1, a clinically relevant kinase target. |
| نوع الوثيقة: |
article in journal/newspaper |
| اللغة: |
unknown |
| العلاقة: |
https://figshare.com/articles/journal_contribution/High-Throughput_Cellular_Thermal_Shift_Assay_Using_Acoustic_Transfer_of_Protein_Lysates/19122797Test |
| DOI: |
10.1021/acschembio.1c00760.s001 |
| الإتاحة: |
https://doi.org/10.1021/acschembio.1c00760.s001Test |
| حقوق: |
CC BY-NC 4.0 |
| رقم الانضمام: |
edsbas.68F4B7B4 |
| قاعدة البيانات: |
BASE |
