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We discuss considerations of promoter reporter selection, assay development and implementation, and image acquisition, analysis, and data handling.
Using the SK-N-MC neuroblastoma cell line a cell based survival selection assay was designed with C2-ceramide or TNFα as an induction signal for apoptosis.
Using this selection assay, we could identify a new single chain antibody binding biotin by growing E. coli BL21 Star (DE3) containing the library in presence of a biotin ampicillin conjugate.
The method was successfully established first for the selection of E. coli BL21 Star (DE3) cells expressing a model single chain antibody (anti-fluorescein) by a simple selection assay on LB-agar plates.
ANAC042 binds to a bipartite cis-regulatory element, as shown by in vitro binding site selection assay (Wu et al., manuscript in preparation).
This result is therefore close to the data obtained independently with the CELD-based binding site selection assay (Wu et al., manuscript in preparation).
Similar(26)
Availability of novel tracers requires dedicated resources and selection assays.
Many of the selection assays are similar to those used for discovery of clinical compounds, although the distribution and clearance of target specific radioligands requires different in vitro and in vivo methods and new derivatives.
Selection assays using 18 trillion cells are infeasible, and multiple sequences introduced into individual cells would compete with each other for loading onto RISC.
Using two different genetic selection assays, we identified constituents within the library that encode small RNAs capable of overcoming both 5-FU and TNF-α-induced apoptosis.
This retrovirally-based expression library was used in two independent unbiased genetic selection assays aimed at overcoming cell death induced by the chemotherapeutic 5-FU or the cytokine TNF-α.
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