OPTIPREP™ APPLICATION 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]
Rhodospirillum rubrum [APPLICATION SHEET S48]
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]
Latest news
Axis-Shield will exhibit at the ASCB meeting in San Francisco December 3-7. You will find us in booth 418
We have recently posted a new package insert for Polymorphprep (see Products in the main menu). In addition to a new figure, we have added a new paragraph; Important practical and sample notes.
We have decided to discontinue the product NycoPrep 1.077, 4x250 ml. We still have lot no. 12GLS12 in stock, with an extended expiry date of November 2017.