To evaluate the different flavonoids as TTR aggregation inhibitors we used a rat Schwannoma (RN22) (American Type Cell Collection) cell line stably transfected with TTR L55P cDNA [22] (link). Cells were grown till 80% cell confluence with the compounds at 1 μM concentration in cell medium (~5 days). Then, cells were incubated for further 24 h still in the presence of compounds but in serum free media. TTR in the medium was quantified by ELISA, and equal amounts of TTR (500 ng) were doted onto a 0.2 μm pore cellulose acetate membrane—dot-blot filter assay. TTR aggregates retained in the membrane were immunodetected using rabbit anti-human TTR (Dako, Glostrup Denmark) (1:500) followed by anti-rabbit HRP antibody (1:1500) and ECL® visualization (GE Healthcare, Buckinghamshire, UK). Quantification of dot-blots was performed with a Bio-Rad ChemiDoc XRS system using the IMAGELAB software. Experiments were repeated at least three times in triplicate.
Rabbit anti human ttr
Manufactured by Agilent Technologies
Sourced in Denmark
Rabbit anti-human TTR is an antibody product used for the detection and quantification of transthyretin (TTR) in human samples. TTR is a transport protein primarily produced in the liver and involved in the transport of thyroid hormones and retinol-binding protein. This antibody can be utilized in various analytical techniques, such as Western blotting, ELISA, and immunohistochemistry, to measure TTR levels in biological samples.
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Lab products found in correlation
2 protocols using rabbit anti human ttr
1
Evaluating Flavonoids as TTR Aggregation Inhibitors
2
Isolation of Human Transthyretin from Plasma
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Human plasma from donors who were informed of the purpose of the study and gave their written consent were collected in accordance with the approved guidelines. Samples were subjected to affinity chromatography to isolate human TTR (hTTR); for this, we used 1 mL column of NHS-activated Sepharose coupled to rabbit anti-human TTR (Dako). The column was washed with PBS and then incubated with 500 μL of human plasma for 2 h at RT. To elute TTR from the column, 5 mL of Gentle Ag/Ab elution buffer (Thermo Scientific) were applied, and 1 mL-aliquots were collected and OD 280 nm was registered.
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