This is a response to a letter by Rubinstein (1).In response to the concerns delineated in a letter by Menachem Rubinstein (1), we found that protein truncation without an endogenous signal peptide is commonly performed in co-immunoprecipitation (2). Furthermore, there was no aggregation or precipitation of misfolded protein in our co-immunoprecipitation assay (Fig. 5B of Ref. 3), and the truncated LGR4-ECD (25–528 amino acids) has well-demonstrated bioactivity, as shown in our previous paper (4). Moreover, the data from our pull-down (Fig. 5C) and Biacore assays (Fig. 5, D and E) confirmed that a strong interaction exists between Lgr4 and intact VSV particles (3).On the question of binding affinity, according to the affinity model, fast estimation of KD is reasonable only when the binding is saturated. In our case, it was hard to reach saturation for every concentration curve. Thus, as in other studies (see, for example, Ref. 5), the kinetics model was the only choice. Upon kinetic regression, the results showed that Kon = 15,450 m−1 s−1, Koff = 0.001067 s−1, and KD = Kd/Ka = 69.05 nm.Finally, the IC50 for LDLR-ECD cited by Dr. Rubinstein refers to sLDLR-mediated inhibition of vesicular stomatitis virus (VSV)–induced cytopathic effects in human WISH cells (Fig. 1A of Ref. 6). In contrast, we detected inhibition of VSV replication by LGR4-ECD in mouse MEF cells by quantitative PCR (Fig. 6A of Ref. 3). It is highly problematic to compare IC50 values determined using different assays, different cells, different expression systems, and different proteins.Taken together, we...
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Publication date: Available online 4 January 2018 Source: European Journal of Radiology Author(s): Peiyao Zhang, Jing Wang, Qin Xu, Zhen...
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Medicine by Alexandros G. Sfakianakis,Anapafseos 5 Agios Nikolaos 72100 Crete Greece,00302841026182, Butyric Acid from Probiotic Staphyloco...
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Does CBD Oil Lower Blood Pressure? This article was originally published at SundayScaries." Madeline Taylor POSTED ON January 13, 20...
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Medicine by Alexandros G. Sfakianakis,Αλέξανδρος Γ. Σφακιανάκης A Novel Technique for Endoscopic Repair of Large Anterior Skull Base Defect...
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2016-11-22T07-55-59Z Source: International Journal of Medical Science and Public Health Banothu Srinivas, Madhu Mohan Reddy B. Backgrou...
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2016-10-15T06-30-01Z Source: The Southeast Asian Journal of Case Report and Review Sangita Deepak Kamath, Neeraj Jain, Saurabh Pathak, Ba...
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BACKGROUND AND PURPOSE: Lesion load is a common biomarker in multiple sclerosis, yet it has historically shown modest association with cl...
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Abstract The development of focused ion beam-scanning electron microscopy (FIB-SEM) techniques has allowed high-resolution 3D imaging of n...
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