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To confirm that TLR8 stimulation of monocytes inhibits matrix degradation we employed a functional matrix breakdown assay.
To investigate whether the TIMP-1 secreted by monocytes is functionally active and impairs substrate breakdown by MMP we employed a functional matrix breakdown assay.
The C1,2C competitive inhibition ELISA assay (a cartilage/skin/bone collagen breakdown assay) measures the carboxy terminus of the primary cleavage site (Col 2 3/4CShort epitope) generated in type I and type II collagens by collagenases [ 11].
We hypothesized that estrogenic EDCs may disrupt progesterone-induced oocyte maturation in the adult amphibian ovary, and tested this with an in vitro germinal vesicle breakdown assay using defolliculated oocytes from the African clawed frog, Xenopus laevis.
For the EC monolayer breakdown assay, 2 × 10 HUVECs were grown to confluent monolayers in fibronectin-coated (10 μg ml−1; Sigma) chamber slides and stimulated with 400 ng CXCL9 for 30 min.
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For the EC monolayer breakdown assays, 2 × 10 human LECs were grown to confluent monolayers and treated with different concentrations of CXCL9 with or without anti-CXCL9 or anti-CXCR3 antibody for 1 h.
Endothelial cell-monolayer breakdown (transmigration) assay was performed using the electrical cell-substrate impedance sensing (ECIS) model 1600R (Applied BioPhysics, Troy, NY, USA).
The C2C competitive inhibition ELISA (a cartilage breakdown-specific assay) measures a related carboxyterminal neoepitope created by the cleavage of only type II collagen by collagenases.
To ensure that these results were not artifacts of global phospholipid depletion or lipid breakdown, we assayed several phosphatidylethanolamines (PtdEtns) not identified as significant by FI-FTICR-MS analysis.
The A. gracilis samples scavenged OH radicals and prevented 2-deoxyribose breakdown in this assay.
Crude methanol extract and derived fractions of J. dolomiaea scavenged OH radicals and prevented 2-deoxyribose breakdown in this assay.
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