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The majority of proteins with nuclear function are actively transported based on its binding to specific karyopherin receptors usually by the presence of NLS and NES signals.
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TFs are families of small proteins that regulate gene expression by binding to specific promoter sequences.
Ran-GTP provides an important source of energy to the reaction by binding to karyopherin β as it enters the nucleus with karyopherin α and cargo in tow, releasing the cargo to the nucleoplasm.
Non-specific binding was subtracted from total binding to obtain specific binding.
Importin, a karyopherin, is involved in the transportation of protein molecules into the nucleus by binding to a specific recognition sequence, called the nuclear localization signal (NLS) [ 46].
It therefore appears that overexpression of Lhp1 from a plasmid interferes with the ability to detect binding to its partner Karyopherin.
Non-specific binding obtained by incubating 1 mM L- glutamate in the binding assay was subtracted from total binding to yield specific [3H] AMPA- binding.
In all experiments, nonspecific binding was determined and subtracted from total binding to obtain specific binding.
Similarly, PY-NLS binding to Karyopherin-β2 (also known as Transportin-1) involves mostly the NLS side chains, and we may observe these NLSs binding to Karyopherin-β2 in the opposite orientation in the future (Lee et al., 2006; Soniat et al., 2013).
Nuclear localization of large proteins, such as LANA, requires a nuclear localization signal (NLS) that mediates binding to members of the karyopherin family of nuclear transport proteins.
At larger probe intensities, non-specific binding becomes less important compared to specific binding.
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