Search

Cell Biology

Categories

    Protocols in Current Issue
    Plasma Membrane Preparation from Lilium davidii and Oryza sativa Mature and Germinated Pollen
    Authors:  Ning Yang, Bing Han, Lingtong Liu, Hao Yang and Tai Wang, date: 05/20/2017, view: 1214, Q&A: 0
    [Abstract] Pollen germination is an excellent process to study cell polarity establishment. During this process, the tip-growing pollen tube will start elongating. The plasma membrane as the selectively permeable barrier that separates the inner and outer cell environment plays crucial roles in this process. This protocol described an efficient aqueous ...
    Isolation of Tonoplast Vesicles from Tomato Fruit Pericarp
    [Abstract] This protocol describes the isolation of tonoplast vesicles from tomato fruit. The vesicles isolated using this procedure are of sufficiently high purity for downstream proteomic analysis whilst remaining transport competent for functional assays. The methodology was used to study the transport of amino acids during tomato fruit ripening (Snowden ...
    Preparation of Golgi Membranes from Rat Liver
    Authors:  Carmen Valente, Gabriele Turacchio, Stefania Spanò, Alberto Luini and Daniela Corda, date: 11/20/2014, view: 2964, Q&A: 0
    [Abstract] This protocol details the isolation of enriched Golgi membranes from rat liver, using discontinuous density gradient centrifugation. This high-yield extraction method is useful for several applications, including immunoprecipitation of solubilised Golgi membrane proteins (preparation included) and electron microscopy. Protocol adapted from ...
    Ciliary and Flagellar Membrane Vesicle (Ectosome) Purification
    Author:  William Dentler, date: 06/20/2014, view: 4022, Q&A: 0
    [Abstract] Eukaryotic cilia/flagella are ideal organelles for the analysis of membrane trafficking, membrane assembly, and the functions of a variety of signal transduction molecules. Cilia are peninsular organelles and the membrane lipids, membrane proteins, and microtubular-associated components are selectively transported into cilia through the region ...
    Purification of HCV-remodeled and Control ER Membranes
    Authors:  David Paul and Ralf Bartenschlager, date: 05/20/2014, view: 2918, Q&A: 0
    [Abstract] As for all positive strand RNA viruses, hepatitis C virus (HCV) RNA replication is tightly associated with rearranged host cell membranes, termed viral replication factories. However, up to now little is known about both viral and cellular constituents of viral replication factories. Here, we describe a protocol to specifically isolate ...
    Preparation of Bacillus subtilis Cell Lysates and Membranes
    Authors:  Juri Niño Bach and Marc Bramkamp, date: 01/20/2014, view: 6521, Q&A: 0
    [Abstract] A common feature of every eukaryotic and prokaryotic cell is that they exhibit a plasma membrane. In Bacillus subtilis (B. subtilis) roughly 25% of all proteins are putative trans- or membrane associated proteins. Here we describe a relatively simple method to separate and prepare membrane and cytosolic proteins by ...
    Membrane Preparation, Sucrose Density Gradients and Two-phase Separation Fractionation from Five-day-old Arabidopsis seedlings
    Authors:  Haibing Yang and Angus Murphy, date: 12/20/2013, view: 9704, Q&A: 0
    [Abstract] Membrane preparation has been widely used for characterization the membrane proteins. Membrane fractions can be separated by a combination of differential and density-gradient centrifugation techniques (Hodges et al., 1972; Leonard and Vanderwoude, 1976). Here we firstly describe a method to isolate total microsomal fractions including ...
    β1 Integrin Cell-surface Immunoprecipitation (Selective Immunoprecipitation)
    Author:  Ralph T. Böttcher, date: 11/05/2013, view: 4590, Q&A: 0
    [Abstract] Immunoprecipitation (IP) is a widely used method to isolate a specific protein from a mixed protein sample using an antibody that exclusively binds to that particular protein. This technique allows studying protein-protein and protein-nucleic acid interactions or to identify post-translational protein modifications. Many proteins, in particular ...
    Isolate and Sub-fractionate Cell Membranes from Caulobacter crescentus
    Author:  Khatira Anwari, date: 10/20/2012, view: 6324, Q&A: 1
    [Abstract] Cell membranes from Caulobacter can be isolated and separated into inner and outer membranes according their characteristic buoyant densities on a sucrose gradient. Fractionation can be used to determine the localisation of uncharacterised proteins and to enrich protein complexes present in either of these membranes for biochemical ...
    Isolation of Inner Membrane Vesicles from Escherichia coli by Using an Affinity Tag
    Authors:  Gang Li and Kevin D. Young, date: 10/20/2012, view: 5811, Q&A: 0
    [Abstract] This protocol was developed in a project aimed to identify the inner membrane proteins localizing to cell poles in Escherichia coli (E. coli). By using a known polar protein Tar as a tag, we isolated pole-derived inner membrane vesicles by affinity capture. The specificity of the polar vesicle isolation was confirmed by mass ...