Iodinated Density Gradient Media

                                             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.


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]



     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 (endosmes/lysosomes/plasma membrane)                  [APPLICATION SHEET S46]

     Lysosome/late endosome events (rat liver)                                         [APPLICATION SHEET S44]

     Methodological review                                                                         [APPLICATION SHEET S42] 

     Self-generated gradients; buoyant density separation                        [APPLICATION SHEET S45]

     Lysosome/late endosome analysis using Nycodenz                           [APPLICATION SHEET S54] 


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]


     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 

Exocyst function

     Plasma membrane domain targeting                                                   [APPLICATION SHEET S47]

     Exocyst vesicle trafficking-methodological summary                           [APPLICATION SHEET S47a]


Exosomes and other microvesicles (mammalian)                               [APPLICATION SHEET S63]

Extracellular vesicles (non-mammalian sources)                                [APPLICATION SHEET S62]

GLUT 4 trafficking (self-generated gradient)                                           [APPLICATION SHEET S48]                                             

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,


Intracellular signalling                                                                             [APPLICATION SHEET S48] 

Light mitochondrial fraction, analysis of

     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”


     Discontinuous gradient                                                                        [APPLICATION SHEET S55]

     Methodological review                                                                         [APPLICATION SHEET S43]

     See also “Light mitochondrial fraction, analysis of”                             

     See also “Endocytosis analysis/Lysosome/late endosome events

Lysosomes (ER/endosomes/plasma membrane)


     Continuous gradient                                                                             [APPLICATION SHEET S56]

Membrane trafficking (endoplasmic reticulum, plasma membranes,

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 S48]

Membrane vesicle/cytosol separation

     See “Protein localization”



     Mammalian (discontinuous gradient)                                                    [APPLICATION SHEET S14]


     Purification and analysis                                                                       [APPLICATION SHEET S14a]                                                                                    

     See also “Light mitochondrial fraction, analysis of”

Neural tissue organelles


      Synaptosomes, synaptoneurosomes, neuromelanin granules and

      synaptic vesicles-a methodological survey                                          [APPLICATION SHEET S40a] 



       Discontinous  gradients and density barrier                                        [APPLICATION SHEET S10]

       Methodological review                                                                        [APPLICATION SHEET S10a] 


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 S64]

Plasma membrane domains


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

     Cationic colloidal silica                                                                         [APPLICATION SHEET S27]

     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 (apical junction complex)”

     NRK cells

        “See Epithelial cells  (apical junction complex)”

     Plasma membrane domain targeting (exocyst)                                    [APPLICATION SHEET S47a]

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 S51]

Secretory granules


     Eosinophils                                                                                           [APPLICATION SHEET S39]

     Pancreatic cells                                                                                    [APPLICATION SHEET S38]

     Adrenal chromaffin, lymphocytic, neutrophil-derived, synaptosomal

     paratid and platelets                                                                             [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




     Organelles various                                                                               [APPLICATION SHEET S59]


     Organelles various                                                                               [APPLICATION SHEET S58]


Arabidopsis thaliana

     See “Plants”


     Extracellular vesicles                                                                           [APPLICATION SHEET S62]

Rhodospirillum rubrum                                                                          


     Organelles, including acidocalsosomes                                               [APPLICATION SHEET S50]

Chlamydomonas reinhardtii                                                                  

     See “Algae”

Dictyostelium discoideum

     See “Protozoa”

Drosophila membranes                                                                          [APPLICATION SHEET S49]


     Extracellular vesicles                                                                           [APPLICATION SHEET S62]

     Organelles                                                                                           [APPLICATION SHEET S59]

     Yeast spheroplast

        Endosomes, ER, Golgi, TGN, vacuole                                             [APPLICATION SHEET S53]

        Membrane trafficking (vacuole, Cvt vesicles etc.)                            [APPLICATION SHEET S52]

        Mitochondria                                                                                     [APPLICATION SHEET S17]

        Peroxisomes                                                                                     [APPLICATION SHEET S57]

Phytomonas francai

      See “Protozoa”

Plant tissues


     Organelles                                                                                            [APPLICATION SHEET S60]

     Proteomic analysis of Arabidopsis                                                       [APPLICATION SHEET S61]

     Protein processing                                                                               [APPLICATION SHEET S61]



     Acidocalcisomes                                                                                  [APPLICATION SHEET S58]

     Contractile vacuoles                                                                            [APPLICATION SHEET S58]

     Other organelles                                                                                  [APPLICATION SHEET S58]

                                                                                                                              OptiPrep Application Sheet Index                                                                                                                                               Subcellular Membranes: February 2020

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