Iodinated Density Gradient Media



♦ 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]


Budded vesicles


     See “Protein localization (membrane versus cytosol)”


     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]


     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,


     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”


     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”



     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,


     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, , 


     See “Plasma membrane/endosomes/TGN/cytosol”

     See “Transport vesicles/endosomes/TGN”




     Acidocalcisomes                                                                                  [APPLICATION SHEET S48]

     Other organelles                                                                                  [APPLICATION SHEET S57]

Amoeba organelles                                                                                [APPLICATION SHEET S56]


Arabidopsis thaliana

     See “Plants”


     Extracellular vesicles                                                                           [APPLICATION SHEET S60]

Rhodospirillum rubrum                                                                          [APPLICATION SHEET S48]

Chlamydomonas reinhardtii                                                                  

     See “Algae”

Dictyostelium discoideum

     See “Protozoa”

Drosophila membranes                                                                          [APPLICATION SHEET S47]


     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]



     Acidocalcisomes                                                                                  [APPLICATION SHEET S48]

     Contractile vacuoles                                                                            [APPLICATION SHEET S48]

     Organelles                                                                                           [APPLICATION SHEET S56]