Protocols in Current Issue
    Measurement of Protein-Protein Interactions through Microscale Thermophoresis (MST)
    Authors:  Magnez Romain, Bryan Thiroux, Morgane Tardy, Bruno Quesnel and Xavier Thuru, date: 04/05/2020, view: 4361, Q&A: 0
    [Abstract] The binding interactions of PD-1 and PD-L1 have been studied by surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) over the past few years, but these investigations resulted in controversy regarding the values of the dissociation constant (Kd) (Freeman et al., 2000). MST is a powerful new method for the ...
    Targeted Nucleotide Substitution in Mammalian Cell by Target-AID
    Authors:  Takayuki Arazoe, Keiji Nishida and Akihiko Kondo, date: 06/05/2017, view: 15798, Q&A: 0
    [Abstract] Programmable RNA-guided nucleases based on CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated protein) systems have been applied to various type of cells as powerful genome editing tools. By using activation-induced cytidine deaminase (AID) in place of the nuclease activity of the CRISPR/Cas9 system, we have ...
    pERK Detection Assays Using the Surefire AlphaScreen® Kit (TGR Biosciences and PerkinElmer)
    Authors:  Cassandra Koole, Denise Wootten and Patrick Sexton, date: 07/05/2013, view: 9419, Q&A: 0
    [Abstract] Extracellular signal-regulated kinase 1 and 2 (ERK1/2) are serine/threonine protein kinases that are phosphorylated on Thr202/Tyr204 (ERK1) and Thr185/Tyr187 (ERK2). Phosphorylation of ERK1/2 (pERK1/2) arises from multiple stimuli, resulting in physiological responses that include cell growth, proliferation and differentiation. This protocol has ...
    cAMP Accumulation Assays Using the AlphaScreen® Kit (PerkinElmer)
    Authors:  Cassandra Koole, Denise Wootten and Patrick Sexton, date: 07/05/2013, view: 11469, Q&A: 0
    [Abstract] Cyclic adenosine monophosphate (cAMP) is an intracellular signaling messenger derived from the catalytic conversion of ATP, and is a major product of activated Gs protein-coupled receptors. Conversely, formation of cAMP is inhibited by Gi protein-coupled receptors. This protocol has been optimized for the detection of ...
    Measurement of Free Cytosolic Calcium Concentration ([Ca2+]i) in Single CHO-K1 Cells
    Authors:  Manuel D. Gahete, Raúl M. Luque and Justo P. Castaño, date: 11/20/2012, view: 12077, Q&A: 0
    [Abstract] This is a protocol to analyze the functional response of single CHO-K1 cells to a given treatment in terms of changes in free cytosolic calcium concentration ([Ca2+]i). This is possible by using the Ca2+ indicator dye Fura-2 AM, a polyamino carboxylic acid that binds to free intracellular calcium and is excited at 340 nm and 380 nm. The ratio of ...

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