Axis-Shield Density Gradient Media

  -a brand of Alere Technologies AS, Oslo, Norway

OPTIPREPAPPLICATION SHEET INDEX

SUBCELLULAR MEMBRANES


♦ The Index is divided into three sections:


A. General methods for preparing gradients; preparing crude organelle or membrane

fractions and analysing the gradient separation


B. An alphabetical subcellular membrane index. Membranes and organelles from

tissues and cells from mammals and most higher eukaryotes are listed according to the

membrane or organelle or subcellular process (e.g. endocytosis).

 

C. Membranes and organelles from protozoa, fungi, algae, plants etc are listed

alphabetically according to the source.


♦ Click on the relevant [APPLICATION SHEET] for a detailed protocol. In some cases

more than one Application Sheet for a specific membrane type may be provided if

different practical strategies are available.


A. GENERAL METHODS


Preparation of gradient solutions (mammalian)                                        [APPLICATION SHEET S01]


Preparation of gradient solutions (non-mammalian)                                 [APPLICATION SHEET S02]


Preparation of discontinuous and continuous gradients                           [APPLICATION SHEET S03]


Preparation of self-generated gradients                                                    [APPLICATION SHEET S04]


Homogenization of mammalian tissues                                                    [APPLICATION SHEET S05]


Homogenization of mammalian cells                                                        [APPLICATION SHEET S06]


Differential centrifugation of homogenates                                               [APPLICATION SHEET S07]


Harvesting gradients                                                                                 [APPLICATION SHEET S08]


Analysis of gradients                                                                                 [APPLICATION SHEET S09]


B. SUBCELLULAR MEMBRANES (MAMMALIAN)


Budded vesicles

 

     See “Protein localization (membrane versus cytosol)”


Caveolae


     See “Plasma membrane domains”


Cytoplasmic vesicles


     See “Protein localization (membrane versus cytosol)”


Cytosol/membrane vesicle separation

 

     See “Protein localization (membrane versus cytosol)”

 

Endocytosis analysis


     Cultured cells: buoyant density gradient analysis                                [APPLICATION SHEET S44]


     Lysosome/late endosome events (rat liver)                                         [APPLICATION SHEET S52]


     Sedimentation velocity gradients                                                         [APPLICATION SHEET S42]


     Self-generated gradients                                                                     [APPLICATION SHEET S43]


Endoplasmic reticulum


     See “Membrane trafficking” for resolution of endoplasmic reticulum from other

     membrane compartments, e.g. plasma membrane, Golgi, trans-Golgi network,

     endosomes, ERGIC


Endoplasmic reticulum domains

 

     Lipid droplets                                                                                       [APPLICATION SHEET S41]


     Mitochondria-associated                                                                     [APPLICATION SHEET S41]


     Perinuclear                                                                                          [APPLICATION SHEET S41]


Endoplasmic reticulum - rough/smooth fractionation


     Cultured cells (continuous gradients)                                                  [APPLICATION SHEET S19]


     Self-generated gradients                                                                     [APPLICATION SHEET S18]


     Separation from Golgi                                                                          [APPLICATION SHEET S20]


Endosomes


     See “Endocytosis analysis”


     See “Membrane trafficking” for resolution of endosomes from other membrane

     compartments, e.g. endoplasmic reticulum, plasma membrane, Golgi, trans-Golgi

     network, ERGIC 


     See “Plasma membrane, TGN/endosomes/cytosol”


     See “Transport vesicles/endosomes/TGN”


Exocyst function


     Plasma membrane domain targeting                                                   [APPLICATION SHEET S45]


Exosomes and other microvesicles                                                      [APPLICATION SHEET S61]


GLUT 4 trafficking


     See “Membrane trafficking”


Golgi and trans-Golgi network


     See “Endoplasmic reticulum, rough/smooth/separation from Golgi”


     See “Membrane trafficking” for resolution of Golgi (or trans-Golgi network) from other

     membrane compartments – e.g. endoplasmic reticulum, plasma membrane, endosomes,


     ERGIC


     See “Plasma membrane, endosomes/TGN/cytosol”


     See “Transport vesicles/endosomes/TGN”


Light mitochondrial fraction, analysis


     Continuous gradient                                                                            [APPLICATION SHEET S15]


     Self-generated gradient                                                                       [APPLICATION SHEET S16]

 

     See also “Lysosomes”


     See also “Mitochondria”


     See also “Peroxisomes”


Lipid rafts


     See “Plasma membrane domains”


Lysosomes


     Discontinuous gradient                                                                        [APPLICATION SHEET S53]


     See also “Light mitochondrial fraction, analysis”


     See also “Endocytosis analysis/Lysosome/late endosome events)


Lysosomes (ER/endosomes/plasma membrane)

 

     Continuous gradient                                                                             [APPLICATION SHEET S54]


Membrane trafficking


     GLUT 4 transport (self-generated gradient)                                         [APPLICATION SHEET S46]


     Intracellular signalling                                                                           [APPLICATION SHEET S46]

 

     Resolution of endoplasmic reticulum and other membrane compartments – e.g. plasma

     membrane, Golgi, trans-Golgi network, endosomes, ERGIC


        Continuous gradient (short spin)                                                       [APPLICATION SHEET S21]


        Continuous gradient (long spin)                                                        [APPLICATION SHEET S22]


        Discontinuous gradient (flotation)                                                      [APPLICATION SHEET S23]


        Discontinuous gradient (sedimentation)                                            [APPLICATION SHEET S24]


        Sedimentation velocity                                                                      [APPLICATION SHEET S25]


        Self-generated gradient                                                                     [APPLICATION SHEET S46]


Membrane vesicle/cytosol separation


     See “Protein localization”


Mitochondria:

 

     Mammalian                                                                                           [APPLICATION SHEET S14]

 

     See also “Light mitochondrial fraction, analysis”


Nuclei                                                                                                        [APPLICATION SHEET S10]


Peroxisomes (mammalian)


     Continuous gradient                                                                              [APPLICATION SHEET S11]


     Discontinuous gradient or density barrier                                             [APPLICATION SHEET S12]


     Self-generated gradient                                                                        [APPLICATION SHEET S13]

 

     See also “Light mitochondrial fraction, analysis”


Plasma membrane


     See “Membrane trafficking” for resolution of plasma membrane from other membrane

     compartments, e.g. endoplasmic reticulum, Golgi, trans-Golgi network, endosomes,


     ERGIC


     Cardiac muscle, from                                                                          [APPLICATION SHEET S26]


     Cationic colloidal silica, using                                                             [APPLICATION SHEET S27]


     Flotation in a discontinuous gradient                                                  [APPLICATION SHEET S62]


Plasma membrane domains

 

     Caco-2 cells (apical/basolateral)                                                          [APPLICATION SHEET S29]


     Caveolae                                                                                              [APPLICATION SHEET S34]


     Epithelial cells (apical junction complex)                                              [APPLICATION SHEET S31]


     Intestinal mucosa (brush border/basolateral)                                       [APPLICATION SHEET S28]

 

     Lipid rafts


        Detergent strategy                                                                             [APPLICATION SHEET S32]


        Detergent-free strategy                                                                      [APPLICATION SHEET S33]


     Renal cortex (brush border/glomeruli/slit diaphragm)                           [APPLICATION SHEET S30]


     MDCK cells


        See “Epithelial cells”


     NRK cells


        “See Epithelial cells”


     Plasma membrane domain targeting (exocyst)                                    [APPLICATION SHEET S45]


Protein localization (large protein complexes)                                     [APPLICATION SHEET S37]

 

Protein localization (membrane versus cytosol)


     Discontinuous flotation gradient                                                           [APPLICATION SHEET S35]


     Self-generated gradient                                                                        [APPLICATION SHEET S36]


Retinal rod outer segments

 

     Optical disks                                                                                         [APPLICATION SHEET S49]


Secretory granules

 

     Eosinophils                                                                                           [APPLICATION SHEET S39]


     Pancreatic cells                                                                                    [APPLICATION SHEET S38]


     Platelets (human)                                                                                 [APPLICATION SHEET S40]


Storage granules (vesicles)


     See “Secretory granules”


Trans-Golgi Network (TGN)


     See “Membrane trafficking” for resolution of trans-Golgi network from other membrane

     compartments, e.g. endoplasmic reticulum, endosomes, plasma membrane, Golgi, , 


     ERGIC


     See “Plasma membrane/endosomes/TGN/cytosol”


     See “Transport vesicles/endosomes/TGN”


C. SUBCELLULAR MEMBRANES (NON-MAMMALIAN)


Algae

 

     Acidocalcisomes                                                                                  [APPLICATION SHEET S48]


     Other organelles                                                                                  [APPLICATION SHEET S57]


Amoeba organelles                                                                                [APPLICATION SHEET S56]

 

Arabidopsis thaliana


     See “Plants”


Bacteria


     Extracellular vesicles                                                                           [APPLICATION SHEET S60]


Chlamydomonas reinhardtii


     See “Algae”


Dictyostelium discoideum


     See “Protozoa”


Drosophila membranes                                                                          [APPLICATION SHEET S47]


Fungi


     Extracellular vesicles                                                                           [APPLICATION SHEET S60]


     Organelles                                                                                           [APPLICATION SHEET S57]


     Yeast spheroplast


        Endosomes, ER, Golgi, TGN, vacuole                                             [APPLICATION SHEET S51]


        Membrane trafficking (vacuole, Cvt vesicles etc.)                            [APPLICATION SHEET S50]


        Mitochondria                                                                                     [APPLICATION SHEET S17]


        Peroxisomes                                                                                     [APPLICATION SHEET S55]


Phytomonas francai


     See “Protozoa”


Plant tissues

 

     Organelles                                                                                            [APPLICATION SHEET S58]


     Proteomic analysis of Arabidopsis                                                       [APPLICATION SHEET S59]


     Protein processing                                                                               [APPLICATION SHEET S58]


Protozoa

 

     Acidocalcisomes                                                                                  [APPLICATION SHEET S48]


     Contractile vacuoles                                                                            [APPLICATION SHEET S48]


     Organelles                                                                                           [APPLICATION SHEET S56]


Rhodospirillum rubrum


     Organelles, including acidocalcisomes                                               [APPLICATION SHEET S48]