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Development and characterization of nanobodies that specifically target the oncogenic Phosphatase of Regenerating Liver-3 (PRL-3) and impact its interaction with a known binding partner, CNNM3

PLoS ONE. 2023-05; 
Caroline N Smith, Kyle Kihn, Zachary A Williamson, K Martin Chow, Louis B Hersh, Konstantin V Korotkov, Daniel Deredge, Jessica S Blackburn
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Proteins, Expression, Isolation and Analysis … Plates were washed three times with 0.05% PBST and loaded with a blocking solution of 0.5… 3 times in PBS and incubated with 1:1000 anti-His HRP antibody (GenScript, A00612, Lot. … Get A Quote

摘要

Phosphatase of Regenerating Liver-3 (PRL-3) is associated with cancer progression and metastasis. The mechanisms that drive PRL-3's oncogenic functions are not well understood, partly due to a lack of research tools available to study this protein. We have begun to address these issues by developing alpaca-derived single domain antibodies, or nanobodies, targeting PRL-3 with a KD of 30-300 nM and no activity towards highly homologous family members PRL-1 and PRL-2. We found that longer and charged N-terminal tags on PRL-3, such as GFP and FLAG, changed PRL-3 localization compared to untagged protein, indicating that the nanobodies may provide new insights into PRL-3 trafficking and function. The nanobodies perf... More

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