Expression and Purification of scFv2H7-P18F3, a Bi-Modular Fusion Protein (BMFP) Targeting Human CD20

    P18F3-based bi-modular fusion proteins (BMFPs), designed to re-direct pre-existing anti-Epstein-Barr virus (EBV) endogenous polyclonal antibodies towards defined target cells, demonstrated efficient biological activity in a mouse tumor model and

    Measuring in vitro ATPase Activity with High Sensitivity Using Radiolabeled ATP
    Authors:  Sarina Veit and Thomas Günther Pomorski, date: 05/20/2023, view: 223, Q&A: 0

    ATPase assays are a common tool for the characterization of purified ATPases. Here, we describe a radioactive [γ-32P]-ATP-based approach, utilizing complex formation with molybdate for phase separation of the free phosphate from

    Determination of Interleukin-17A and Interferon-γ Production in γδ, CD4+, and CD8+ T Cells Isolated from Murine Lymphoid Organs, Perivascular Adipose Tissue, Kidney, and Lung
    Authors:  Kevin Comeau, Antoine Caillon, Pierre Paradis and Ernesto L. Schiffrin, date: 05/20/2023, view: 203, Q&A: 0

    T cells localized to the kidneys and vasculature/perivascular adipose tissue (PVAT) play an important role in hypertension and vascular injury. CD4+, CD8+, and γδ T-cell subtypes are programmed to produce interleukin (IL)-17 or

    A Simple, Reproducible Procedure for Chemiluminescent Western Blot Quantification
    Authors:  Daniel Cañada-García and Juan C. Arévalo, date: 05/05/2023, view: 668, Q&A: 0

    Western blotting is a universally used technique to identify specific proteins from a heterogeneous and complex mixture. However, there is no clear and common procedure to quantify the results obtained, resulting in variations due to the

    A Modified Acyl-RAC Method of Isolating Retinal Palmitoyl Proteome for Subsequent Detection through LC-MS/MS

    Palmitoylation is a unique and reversible posttranslational lipid modification (PTM) that plays a critical role in many cellular events, including protein stability, activity, membrane association, and protein–protein interactions. The dynamic

    In vitro Reconstitution of Phase-separated p62 Bodies on the Arp2/3-derived Actin Network
    Authors:  Tong Liu, Mengbo Xu and Na Mi, date: 04/20/2023, view: 322, Q&A: 0

    In cells, p62/SQSTM1 undergoes liquid–liquid phase separation (LLPS) with poly-ubiquitin chains to form p62 bodies that work as a hub for various cellular events, including selective autophagy. Cytoskeleton components such as Arp2/3-derived

    Determination of Paraquat in Arabidopsis Tissues and Protoplasts by UHPLC-MS/MS
    Authors:  Mingming Zhao, Qi Wang, Muyu Shi, Ziyan Sun, Huiru Tang and Xiaochun Ge, date: 04/05/2023, view: 318, Q&A: 0

    Paraquat is a cost-effective herbicide, widely used in many countries, that can induce severe oxidative stress in photosynthetic tissues. Studying plant herbicide resistance or antioxidant stress mechanisms requires determining the cellular paraquat

    Rapid and Reliable Quantification of Glycerol-3-phosphate Using Gas Chromatography–coupled Mass Spectrometry
    Authors:  Huazhen Liu, Keshun Yu and Pradeep Kachroo, date: 04/05/2023, view: 287, Q&A: 0

    Glycerol-3-phosphate (G3P) is a conserved precursor of glycerolipids that also plays an important role in plant defense. Its levels and/or metabolism are also associated with many human disorders including insulin resistance, diabetes, obesity, and

    Isolation of Immunocomplexes from Zebrafish Brain
    Authors:  Jennifer Carlisle Michel and Adam C. Miller, date: 04/05/2023, view: 188, Q&A: 0

    Zebrafish is an excellent model to study vertebrate neurobiology, but its synaptic components that mediate and regulate fast electrical synaptic transmission are largely unidentified. Here, we describe methods to solubilize and immunoprecipitate

    Separating Inner and Outer Membranes of Escherichia coli by EDTA-free Sucrose Gradient Centrifugation
    Authors:  Sheng Shu and Wei Mi, date: 03/20/2023, view: 422, Q&A: 0

    The envelope of Gram-negative bacteria consists of an outer membrane (OM), a peptidoglycan cell wall, and an inner membrane (IM). The OM and IM have different components of proteins and lipids. Separating the IM and OM is a basic biochemical ...

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