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Thermal sensitivity was evaluated using either a reflexive measure of hindpaw withdrawal latency to a radiant heat source or using an orofacial operant thermal assay.
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No group differences were found at baseline for both mechanical and thermal assays.
The worms for thermal stress assay, oxidative stress assay and paralysis assay were fed with 10 µL of stock solution in the OP50 food source, seeded on NGM plates and starting from egg.
Thermal shift assay: Thermal shift assays were performed on an Mx3005P quantitative PCR instrument (Stratagene, La Jolla, California) capable of temperature control and fluorescence detection.
In vivo thermal shift assay (cellular thermal shift assay, CTSA): HepG2 cells (3 × 107) were seeded into a 100-mm culture plate with 12 μmol/L of G-Rg5 or G-Rk1 and cultured for 1 h.
Fluorescence based thermal aggregation assay, in vitro protection assay via IPP at various temperatures and ELISA confirmed that NTD125 displayed chaperone-like function in stabilizing the wild type conformation and restoring the mutant phenotype at elevated temperature.
Thermal denaturation assay: The thermal-shift denaturation assay was performed on an iCycler iQ Real Time Detection System (Bio-Rad) in 96-well iCycler iQ PCR plates sealed with optically clear lids.
We have resorted to the thermal shift assay methodology to assess the predictions from VLS [13].
Figure 1 Flowchart of the overall approach and the thermal shift assay results.
In contrast, the thermal shift assay can accurately determine picomolar affinities.
(B) Cellular thermal shift assay presented the thermal stability of Annexin A2 under treatment with G-Rg5 (12 μmol/L) and G-Rk1 (12 μmol/L).
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