The in vitro pull-down assay is a well-established method to confirm direct binding in protein-protein interactions that was inferred from other interaction assays, such as two-hybrid analysis (1). The assay is usually carried out using glutathione-S-transfer ase-tagged or His-tagged fusion pro...
This was further supported by in vitro pull-down assays, in which bacterially produced UPF1 was observed to associate with SMG5/7 and SMG6 in a manner enhanced by phosphorylation with recombinant SQ-specific ATM kinase (Fig. 5d). Collectively, the observations in Fig. 5 demonstrate UPF1 ...
The presence of an endogenous interaction and the results of the in vitro pull-down assays revealed that DC-SIGN binds directly with TLR4. We also present evidence that DC-SIGN mediates TLR4-regulated NFκB activation but not activation of p38 and JNK. Our results suggest an essential role of...
f-i. Transwell assays were employed to evaluate the impact of SNAI1 overexpression or suppression on the in vitro migration and invasion ability of TET cells. Representative micrographs and quantification of cells attached to the lower surface of the chamber are presented (Scale bars: 200 μm)....
Some research groups have tried to study the transcriptome of living cells in vitro. For example, TIVA has pioneered the capture of mRNA in living cells for transcriptional analysis [42]. However, this method is currently not applicable to the analysis of many cells. ZipSeq marks DNA codes (...
East Carolina University. Commonly used, highly accessible methods for examining cell migration and invasion in vitro are described. The first method is the cell wound closure assay that measures cell motility. The second method is the transwell migrati
in vitro stimulation of EphA2 signaling, Immunoprecipitation, Flow Cytometry, Functional Assays, Agonistic Antibodies Sakamoto A, Kato K, Hasegawa T, Ikeda S. (2018). "An Agonistic Antibody to EPHA2 Exhibits Antitumor Effects on Human Melanoma Cells" Anticancer Res 38(6):3273-3282. ...
To test this possibility, we performed in vitro pull-down assays using purified OsSRF3-His, UgsL-MBP, and OsBAK1-GST fusion proteins. When we incubated the equal amounts of OsSRF3-His and OsBAK1-GST proteins with GST beads and added increasing amounts of UgsL protein to the OsBAK1–...
To better understand how Region 2 mutations alter NEAT1 function, and evaluate if mutation could affect the binding of proteins to NEAT1, we compared the protein interactome of wild-type and mutant RNA by in vitro pulldown coupled to mass spectrometry (Fig. 6d). We created a 288 nt fragmen...
Cell migration and invasion have essential roles in both normal physiology and disease. As such, methodologies to assess cell migratory and invasive capacities are necessary to elucidate normal cell processes and underlying mechanisms of disease. Here, we describe commonly used transwell in vitro methods...