Mertk

Manufactured by Sino Biological

The MerTK is a laboratory protein that functions as a receptor tyrosine kinase. It plays a role in the regulation of cell growth, survival, and phagocytosis.

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Lab products found in correlation

Spin desalting column, by Thermo Fisher Scientific (2 mentions) Galectin 3, by Merck Group (2 mentions) Pherastar plus, by BMG Labtech (2 mentions) Alexa fluor 488 nhs ester, by Thermo Fisher Scientific (1 mentions)

2 protocols using mertk

Galectin-3 Binding Kinetics Assay

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Galectin-3 (Sigma-Aldrich) was labeled with Alexa Fluor 488 NHS Ester (succinimidyl ester) in PBS at room temperature for 30 min, and free dye was washed with desalting spin columns (Thermo Scientific). Labeling was checked by MALDI mass spectrometry. Fluorescence anisotropy measurements were recorded using a PHERAstar Plus multidetection plate reader (BMG Labtech) equipped with a fluorescence polarization optic module (λex = 485 nm, λem = 520 nm) at 25°C. Each data point is the mean of 200 flashes per well. The voltage gain was set by adjusting the target mP values of labeled Gal-3 to that of fluorescein (35 mP). Serial dilutions of MerTK (Sino Biological), modified citrus pectin (ecoNugenics), and TREM2 ectodomain were made in PBS and added together in a 1:1 ratio to a constant concentration of Gal-3 (35 nM; 44 mP). K_d was estimated, assuming a 1:1 binding, using GraphPad Prism. Purified TREM2 ectodomain was a kind gift from Roger Dodd and Peter St. George-Hyslop (University of Cambridge).

Nomura K., Vilalta A., Allendorf D.H., Hornik T.C, & Brown G.C. (2017). Activated Microglia Desialylate and Phagocytose Cells via Neuraminidase, Galectin-3, and Mer Tyrosine Kinase. The Journal of Immunology Author Choice, 198(12), 4792-4801.

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Galectin-3 Binding Affinity Assay

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Galectin 3 (Sigma) was labelled with Alexa Fluor® 488 NHS Ester (Succinimidyl Ester) in PBS at RT for 30 minutes and free dye was washed with desalting spin columns (Thermo Scientific). Labelling was checked by MALDI mass spectrometry. Fluorescence anisotropy measurements were recorded in a PHERAstar Plus multi-detection plate reader (BMG Labtech) equipped with a fluorescence polarization optic module (λex = 485 nm, λem = 520 nm) at 25°C. Each data point is the mean of 200 flashes per well. The voltage gain was set by adjusting the target mP values of labelled Galectin 3 to that of fluoresceine (35 mP). Serial dilutions of MerTK (Sino Biological Inc), modified citrus pectin (ecoNugenics) and TREM2 ectodomain were made in PBS and added together in 1:1 ratio to a constant concentration of galectin-3 (35nM; 44mP). The K_d was estimated assuming a one-to-one binding using GraphPad Prism. Purified TREM2 ectodomain was a kind gift of Roger Dodd and Peter St George-Hyslop (Cambridge).

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