scholarly journals Tissue spheroid fusion-based in vitro screening assays for analysis of tissue maturation

2010 ◽  
Vol 4 (8) ◽  
pp. 659-664 ◽  
Author(s):  
Zoltan Hajdu ◽  
Vladimir Mironov ◽  
Agnes Nagy Mehesz ◽  
Russell A. Norris ◽  
Roger R. Markwald ◽  
...  
Keyword(s):  
In Vitro Screening ◽  
Screening Assays

scholarly journals Preparative Scale Production of Recombinant Human Transthyretin for Biophysical Studies of Protein-Ligand and Protein-Protein Interactions

2020 ◽  
Vol 21 (24) ◽  
pp. 9640
Author(s):  
Ellen Y. Cotrina ◽  
Marta Vilà ◽  
Joan Nieto ◽  
Gemma Arsequell ◽  
Antoni Planas
Keyword(s):  
Binding Protein ◽  
Amyloid Β ◽  
Retinol Binding Protein ◽  
Familial Amyloidotic Polyneuropathy ◽  
Main Role ◽  
Scale Production ◽  
In Vitro Screening ◽  
Preparative Scale ◽  
Screening Assays

Human transthyretin (hTTR), a serum protein with a main role in transporting thyroid hormones and retinol through binding to the retinol-binding protein, is an amyloidogenic protein involved in familial amyloidotic polyneuropathy (FAP), familial amyloidotic cardiomyopathy, and central nervous system selective amyloidosis. hTTR also has a neuroprotective role in Alzheimer disease, being the major Aβ binding protein in human cerebrospinal fluid (CSF) that prevents amyloid-β (Aβ) aggregation with consequent abrogation of toxicity. Here we report an optimized preparative expression and purification protocol of hTTR (wt and amyloidogenic mutants) for in vitro screening assays of TTR ligands acting as amyloidogenesis inhibitors or acting as molecular chaperones to enhance the TTR:Aβ interaction. Preparative yields were up to 660 mg of homogenous protein per L of culture in fed-batch bioreactor. The recombinant wt protein is mainly unmodified at Cys10, the single cysteine in the protein sequence, whereas the highly amyloidogenic Y78F variant renders mainly the S-glutathionated form, which has essentially the same amyloidogenic behavior than the reduced protein with free Cys10. The TTR production protocol has shown inter-batch reproducibility of expression and protein quality for in vitro screening assays.

Getting a handle on chemical probes of chomatin readers

2020 ◽  
Author(s):  
Jarod M Waybright ◽  
Lindsey I James
Keyword(s):  
Target Localization ◽  
In Vitro Screening ◽  
Chemical Probes ◽  
Dynamic Nature ◽  
Post Translational Modifications ◽  
Epigenetic Machinery ◽  
Cellular Target ◽  
Screening Assays ◽  
A Site

The dynamic nature of histone post-translational modifications such as methylation or acetylation makes possible the alteration of disease associated epigenetic states through the manipulation of the associated epigenetic machinery. One approach is through small molecule perturbation. Chemical probes of epigenetic reader domains have been critical in improving our understanding of the biological consequences of modulating their targets, while also enabling the development of novel probe-based reagents. By appending a functional handle to a reader domain probe, a chemical toolbox of reagents can be created to facilitate chemiprecipitation of epigenetic complexes, evaluate probe selectivity, develop in vitro screening assays, visualize cellular target localization, enable target degradation and recruit epigenetic machinery to a site within the genome in a highly controlled fashion.

In Vitro Screening of Azalea for Resistance to Azalea Lace Bug

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 506b-506
Author(s):  
Carol D. Robacker ◽  
S.K. Braman
Keyword(s):  
Leaf Damage ◽  
In Vitro Screening ◽  
Screening Assay ◽  
Delaware Valley ◽  
In Vitro Techniques ◽  
Culture Tube ◽  
Lace Bug ◽  
Lace Bugs ◽  
Laboratory Bioassays

Azalea lace bug (Stephanitis pyrioides) is the most serious pest on azalea. Results of laboratory bioassays and field evaluations of 17 deciduous azalea taxa have identified three resistant taxa: R. canescens, R. periclymenoides, and R. prunifolium. Highly susceptible taxa are `Buttercup', `My Mary', R. oblongifolium, and the evergreen cultivar `Delaware Valley White'. To determine whether in vitro techniques would have potential value in screening or selecting for resistance, or for the identification of morphological or chemical factors related to resistance, an in-vitro screening assay was developed. In-vitro shoot proliferation was obtained using the medium and procedures of Economou and Read (1984). Shoots used in the bioassays were grown in culture tubes. Two assays were developed: one for nymphs and one for adult lace bugs. To assay for resistance to nymphs, `Delaware Valley White' leaves containing lace bug eggs were disinfested with 70% alcohol and 20% commercial bleach, and incubated in sterile petri plates with moistened filter paper until the nymphs hatched. Five nymphs were placed in each culture tube, and cultures were incubated for about 2 weeks, or until adults were observed. To assay for resistance to adults, five female lace bugs were placed in each culture tube and allowed to feed for 5 days. Data collected on survival and leaf damage was generally supportive of laboratory bioassays and field results. Adult lace bugs had a low rate of survival on resistant taxa. Survival of nymphs was somewhat reduced on resistant taxa.

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