A single-step method for rapid extraction of total lipids from green microalgae.
Microalgae produce a wide range of lipid compounds of potential commercial interest. Total lipid extraction performed by conventional extraction methods, relying on the chloroform-methanol solvent system are too laborious and time consuming for screening large numbers of samples. In this study, three previous extraction methods devised by Folch et al. (1957), Bligh and Dyer (1959) and Selstam and Öquist (1985) were compared and a faster single-step procedure was developed for extraction of total lipids from green microalgae. In the single-step procedure, 8 ml of a 2∶1 chloroform-methanol (v/v) mixture was added to fresh or frozen microalgal paste or pulverized dry algal biomass contained in a glass centrifuge tube.
The biomass was manually suspended by vigorously shaking the tube for a few seconds and 2 ml of a 0.73% NaCl water solution was added. Phase separation was facilitated by 2 min of centrifugation at 350 g and the lower phase was recovered for analysis. An uncharacterized microalgal polyculture and the green microalgae Scenedesmus dimorphus, Selenastrum minutum, and Chlorella protothecoides were subjected to the different extraction methods and various techniques of biomass homogenization. The less labour intensive single-step procedure presented here allowed simultaneous recovery of total lipid extracts from multiple samples of Centrifuge tube green microalgae with quantitative yields and fatty acid profiles comparable to those of the previous methods. While the single-step procedure is highly correlated in lipid extractability (r² = 0.985) to the previous method of Folch et al. (1957), it allowed at least five times higher sample throughput.
Santowhite, a commercial antioxidant used in the manufacture of polypropylene, contaminates 12-ml polyallomer tubes to the extent of 0.2-0.3 mg/tube.
- It is distributed through the plastic and appears as a microscopic dust on the tubes’ surfaces. In the preparation of polysomes from rat liver by standard methods, approximately 15% of the antioxidant in previously water-washed tubes was removed by suspension of the polysome pellet with stirring in 1-2 ml of Tris buffer, pH 7.8.
- The polyallomer impurity has been shown to be identical with Santowhite, which is 4,4′-butylidene-bis(6-tert-butyl-m-cresol), by UV, mass, and NMR spectra.
- It is not uniformly removed from polyallomer tubes by common detergents but is removed by washing with acetone, to which the plastic is resistant.
- Colloidal silica–aluminum modified–PVP density gradient centrifugation: centrifuge tube wall cell adherence, aggregation, separation properties and comparison to BSA and Ficoll.
A method is described for the inexpensive and easy preparation of colloidal silica–aluminum modified–polyvinylpyrrolidone (CS-AM-PVP) density gradient centrifugation medium.
Using density gradient centrifugation, several cell separation and biochemical characteristics were studied: centrifuge tube wall cell adherence, mouse spleen cell density distribution, rebanding properties, mitogen response and cell aggregation. Cell adherence to the centrifuge tube wall using density gradients of CS-AM-PVP was compared with density gradients of bovine serum albumin and Ficoll.
Few cells adhered to the centrifuge tube wall when CS-AM-PVP was used as a gradient medium; whereas, significant cell adherence to the centrifuge tube cell wall occurred when bovine serum albumin or Ficoll was used as a gradient medium. The CS-AM-PVP gradient medium did not inhibit the response to mitogens of mouse spleen cells which had been separated into density subpopulations in a discontinuous CS-AM-PVP density gradient, caused a minimum amount of cell aggregation, and was found to be non-toxic.
Islet transplantation is a promising method for restoring normoglycemia and alleviating the long term complications of diabetes.
- Widespread application of islet transplantation is hindered by the limited supply of human islets and requires a large increase in the availability of suitable insulin secreting tissue as well as robust quality assessment methodologies that can ensure safety and in vivo efficacy.
- We explore the application of nuclear magnetic resonance (NMR) spectroscopy in two areas relevant to beta cell engineering and islet transplantation: (1) the effect of genetic alterations on glucose metabolism, and (2) quality assessment of islet preparations prior to transplantation.
- Results obtained utilizing a variety of NMR techniques demonstrate the following: (1) Transfection of Rat1 cells with the c-myc oncogene (which may be involved in cell proliferation and cell cycle regulation) and overexpression of Bcl-2 (which may protect cells from stresses such as hypoxia and exposure to cytokines) introduce a wide array of alterations in cellular biochemistry, including changes in anaerobic and oxidative glucose metabolism, as assessed by 13C and 31P NMR spectroscopy.
- Overnight incubation of islets and beta cells in the bottom of centrifuge tubes filled with medium at room temperature, as is sometimes done in islet transportation, exposes them to severe oxygen limitations that may cause cell damage. Such exposure, leading to reversible or irreversible damage, can be observed with NMR-detectable markers using conventional 13C and 31P NMR spectroscopy of extracts.
- In addition, markers of irreversible damage (as well as markers of hypoxia) can be detected and quantified without cell extraction using high-resolution magic angle spinning 1H NMR spectroscopy. Finally, acute ischemia in a bed of perfused beta cells leads to completely reversible changes that can be followed in real time with 31P NMR spectroscopy
Plasma membranes were isolated from rat pheochromocytoma cells (PC-12) grown in spinner culture.
The rapid and simple isolation procedure consisted of a differential and isopycnic centrifugation (in a linear sucrose gradient) with the aid of a high capacity fixed angle rotor equipped with siliconized centrifuge tubes. The isolated membranes were closed and osmotically active vesicles (about 0.3 micron in diameter) with a mean intravesicular water space of 1.84 microliters/mg protein. In the presence of an inward gradient of sodium chloride and an outward gradient of potassium, [3H]noradrenaline (50 nM) was taken up and accumulated 550-fold (at 31 degrees C).
Centrifuge Tubes, 50 ml , Black ,PP,Sterile |
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LC8008-025 | GenDepot | 25/Bag | 100.8 EUR |
Centrifuge Tubes, 50 ml , Black ,PP,Sterile |
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LC8008-500 | GenDepot | 500/CS | 586.8 EUR |
Centrifuge Tubes, 50 ml ,PP,Sterile,Plug-Seal Cap, in Bag |
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LC8005-500 | GenDepot | 500/CS | 241.2 EUR |
Centrifuge Tubes, 15 ml , Black ,PP,Sterile |
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LC8007-050 | GenDepot | 50/Bag | 121.2 EUR |
Centrifuge Tubes, 15 ml , Black ,PP,Sterile |
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LC8007-500 | GenDepot | 500/CS | 440.4 EUR |
Centrifuge Tubes, 15 ml ,PP,Sterile,Plug-Seal Cap, in Bag |
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LC8003-500 | GenDepot | 500/CS | 208.8 EUR |
Centrifuge Tubes, 50 ml ,PP,Sterile,Plug-Seal Cap,Self Standing, in Bag |
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LC8006-500 | GenDepot | 500/CS | 360 EUR |
Corning LSE Single Block 5 x 50 mL Centrifuge Tubes |
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480129 | Scientific Laboratory Supplies | EACH | 117.42 EUR |
Test Tube rack for 15 x 50 ml tubes |
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 -B2000-4-T500 | Benchmark Scientific | 1 PC | 252.35 EUR |
Test Tube rack for 15 x 50 ml tubes |
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B2000-4-T500 | Benchmark Scientific | 1 each | 109.3 EUR |
LV CryoOil 50 ml |
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LVCO-50 | MiTeGen | 50 ml | 245 EUR |
Centrifuge Tubes, 50 ml ,PP,Sterile,Plug-Seal Cap, in Rack |
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LC8004-500 | GenDepot | 500/CS | 267.6 EUR |
29 mm Test Tube Foam Rack (for 50 mL centrifuge tubes) |
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MIX1118 | Scientific Laboratory Supplies | EACH | 42.18 EUR |
Box for 16 tubes 50 mL |
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DD45548 | Scientific Laboratory Supplies | PK2 | 79.8 EUR |
Eppendorf Adapter for 0.5 mL micro- centrifuge tubes + 0.6 ml Microtainer set of 6 |
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E5425716001 | Scientific Laboratory Supplies | PK6 | 14.76 EUR |
Centrifuge Tubes, 15 ml ,PP,Sterile,Plug-Seal Cap, in Rack |
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LC8002-500 | GenDepot | 500/CS | 218.4 EUR |
16 mm Test Tube Foam Rack (for 15 mL centrifuge tubes) |
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MIX1098 | Scientific Laboratory Supplies | EACH | 42.18 EUR |
ERLENMEYER FLASK, 50 ML, PP |
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4985P-50 | CORNING | 6/pk | 60 EUR |
ERLENMEYER FLASK, 50 ML, PMP |
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4990P-50 | CORNING | 6/pk | 64.8 EUR |
Platform for 4 x 50 mL tubes |
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MIX1254 | Scientific Laboratory Supplies | EACH | 47.88 EUR |
CoolRack 50 ml 4 x 50ml tubes |
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432062 | Scientific Laboratory Supplies | EACH | 359.1 EUR |
CENTRIFUGE TUBE 50 ML 30 X 115 MM |
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T2318-500EA | Scientific Laboratory Supplies | PK500 | 223.44 EUR |
Platform for 8 x 15 ml tubes |
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MIX1260 | Scientific Laboratory Supplies | EACH | 47.88 EUR |
Adapter 8 x 15 mL Conical Tubes |
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E5804783000 | Scientific Laboratory Supplies | EACH | 182.4 EUR |
Cardboard cryobox for 36 tubes 15 mL |
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DD10194 | Scientific Laboratory Supplies | EACH | 13.68 EUR |
Block, 15 x1.5ml centrifuge tubes |
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BCM1405-B15 | Bio Basic | 1 pcs, 1 UNIT | 194.76 EUR |
0.8 ml Storage Plate (1 bag 50 plates) |
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AB-0859 | Scientific Laboratory Supplies | PK50 | 274.74 EUR |
FlexiRack Tube Insert for 50 mL Tubes-Orange |
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ARG1754 | Scientific Laboratory Supplies | PK5 | 11.27 EUR |
White polypropylene rack for 18 tubes 50 mL |
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DD39180042 | Scientific Laboratory Supplies | EACH | 25.08 EUR |
Syringe Injekt 2 pieces volume 2 mL |
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DD921001 | Scientific Laboratory Supplies | PK100 | 10.26 EUR |
Strip Of 4 0.1 Ml PCR Tubes |
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PCR1188 | Scientific Laboratory Supplies | PK250 | 132.24 EUR |
Rack for 64 tubes of 1.5 mL |
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DD764046 | Scientific Laboratory Supplies | EACH | 12.2 EUR |
Naked Gold 40 nm Concentrated Gold Sol, 50 O.D./mL, 1 mL |
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NGL40-B001 | BioAssayWorks | 1 mL | 287.09 EUR |
Naked Gold 40 nm Concentrated Gold Sol, 50 O.D./mL, 9 mL |
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NGL40-B009 | BioAssayWorks | 9 mL | 1424.16 EUR |
Naked Gold 40 nm Concentrated Gold Sol, 50 O.D./mL, 44 mL |
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NGL40-B044 | BioAssayWorks | 44 mL | 5336.71 EUR |
Naked Gold 40 nm Concentrated Gold Sol, 50 O.D./mL, 100 mL |
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NGL40-B100 | BioAssayWorks | 100 mL | 10165.37 EUR |
Naked Gold 40 nm Concentrated Gold Sol, 15 O.D./mL, 1 mL |
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NG40-B001 | BioAssayWorks | 1 mL | 128.09 EUR |
Naked Gold 40 nm Concentrated Gold Sol, 15 O.D./mL, 9 mL |
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NG40-B009 | BioAssayWorks | 9 mL | 468.78 EUR |
Naked Gold 40 nm Concentrated Gold Sol, 15 O.D./mL, 44 mL |
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NG40-B044 | BioAssayWorks | 44 mL | 1640.08 EUR |
Naked Gold 40 nm Concentrated Gold Sol, 15 O.D./mL, 100 mL |
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NG40-B100 | BioAssayWorks | 100 mL | 3088.55 EUR |
Test Tube rack for 40 x 0.5 ml tubes |
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B2000-4-T5 | Benchmark Scientific | 1 PC | 153.17 EUR |
0.2 ml 8-Strip Tubes, 120 Strips of 8 Tubes |
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GP301 | GenDepot | 960 tubes | 166.8 EUR |
0.2 ml 8-Strip Tubes, 250 Strips of 8 Tubes |
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GP302 | GenDepot | 2,000 tubes | 380.4 EUR |
0.2 ml 12-Strip Tubes, 80 Strips of 12 Tubes |
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GP305 | GenDepot | 960 tubes | 295.2 EUR |
Test Tube rack for 30 x 50 ml tubes |
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  B2000-8-T500 | Benchmark Scientific | 1 PC | 2995.98 EUR |
Test Tube rack for 30 x 50 ml tubes |
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B2000-8-T500 | Benchmark Scientific | 1 each | 219.7 EUR |
Bottle Centrifuge Pp 250 Ml |
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3141-0250 | Scientific Laboratory Supplies | PK4 | 116.28 EUR |
Bottle Centrifuge Pp 450 Ml |
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3141-0500 | Scientific Laboratory Supplies | PK4 | 152.76 EUR |
0.2 ml 8-Strip Tubes & Flat Caps, 250 Strips of 8 Tubes and Caps |
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GP203 | GenDepot | 2000 tubes/caps | 333.6 EUR |
0.2 ml 8-Strip Tubes & Domed Caps, 120 Strips of 8 Tubes and Caps |
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GP204 | GenDepot | 960 tubes/caps | 248.4 EUR |
0.2 ml 8-Strip Tubes & Domed Caps, 250 Strips of 8 Tubes and Caps |
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GP205 | GenDepot | 2,000 tubes/caps | 348 EUR |
0.3 ml 8-Strip Tubes & Domed Caps, 250 Strips of 8 Tubes and Caps |
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GP209 | GenDepot | 2,000 tubes/caps | 400.8 EUR |
0.2 ml 12-Strip Tubes & Flat Caps, 80 Strips of 12 Tubes and Caps |
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GP210 | GenDepot | 960 tubes/caps | 319.2 EUR |
0.2 ml 12-Strip Tubes & Domed Caps, 80 Strips of 12 Tubes and Caps |
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GP211 | GenDepot | 960 tubes/caps | 319.2 EUR |
0.2 ml 8-Strip Tubes & Flat Caps, 125 Strips of 8 Tubes and Caps |
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GP903 | GenDepot | 1000 tubes/caps | 241.2 EUR |
0.2 ml 8-Strip Tubes & Dome Caps, 125 Strips of 8 Tubes and Caps |
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GP904 | GenDepot | 1000 tubes/caps | 241.2 EUR |
50 ml Reagent Reservoir |
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GR238016 | Genorise Scientific | Pack of 200 | 177.6 EUR |
TALON SUPERFLOW 50 ML |
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GE28-9575-02 | Scientific Laboratory Supplies | EACH | 727.32 EUR |
Reagent reservoir 50 mL |
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DD53482 | Scientific Laboratory Supplies | PK100 | 194.94 EUR |
SOURCE 15S 50 mL |
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17094401 | Scientific Laboratory Supplies | EACH | 808.26 EUR |
SOURCE 15Q 50 mL |
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17094701 | Scientific Laboratory Supplies | EACH | 808.26 EUR |
SOURCE 30S 50 mL |
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17127301 | Scientific Laboratory Supplies | EACH | 835.62 EUR |
SOURCE 30Q 50 mL |
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17127501 | Scientific Laboratory Supplies | EACH | 835.62 EUR |
TALON Superflow 50 ml |
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28957502 | Scientific Laboratory Supplies | EACH | 810.54 EUR |
Test Tube rack for 41 x 15 ml tubes |
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  B2000-4-T150 | Benchmark Scientific | 1 PC | 164.65 EUR |
Test Tube rack for 76 x 15 ml tubes |
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  B2000-8-T150 | Benchmark Scientific | 1 PC | 252.35 EUR |
Test Tube rack for 41 x 15 ml tubes |
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B2000-4-T150 | Benchmark Scientific | 1 each | 109.3 EUR |
Test Tube rack for 76 x 15 ml tubes |
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B2000-8-T150 | Benchmark Scientific | 1 each | 219.7 EUR |
50ml Conical Centrifuge Tubes, Green Cap, 25/Bag |
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CT788-G | Bio Basic | 1PK, 25UNIT | 67.31 EUR |
50ml Conical Centrifuge Tubes, Yellow Cap, 25/Bag |
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CT788-Y | Bio Basic | 1PK, 25UNIT | 67.31 EUR |
ML-297 |
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C5045-50 | ApexBio | 50 mg | 1245.6 EUR |
ML-090 |
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C4746-50 | ApexBio | 50 mg | 216 EUR |
ML-335 |
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C4792-50 | ApexBio | 50 mg | 907.2 EUR |
ML 228 |
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A4508-50 | ApexBio | 50 mg | 1417.2 EUR |
ML-204 |
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9448-50 | Biovision | 764.4 EUR | |
ML 348 |
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B4933-50 | ApexBio | 50 mg | 1216.8 EUR |
ML 213 |
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B5628-50 | ApexBio | 50 mg | 793.2 EUR |
ML 171 |
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B5672-50 | ApexBio | 50 mg | 150 EUR |
ML 204 |
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B5696-50 | ApexBio | 50 mg | 986.4 EUR |
ML SA1 |
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B5700-50 | ApexBio | 50 mg | 954 EUR |
ML 190 |
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B5741-50 | ApexBio | 50 mg | 1746 EUR |
ML 337 |
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B5786-50 | ApexBio | 50 mg | 1616.4 EUR |
ML 145 |
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B7604-50 | ApexBio | 50 mg | 1726.8 EUR |
ML 154 |
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B7765-50 | ApexBio | 50 mg | 2018.4 EUR |
LCP EZ-Cut films (50 pieces, with liners) |
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MLCP-EZCTF-50 | MiTeGen | 50 FILMS | 234 EUR |
0.2 ml 8-Strip Tubes & Optically Clear Caps, 120 Strips of 8 Tubes and Caps |
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GP207 | GenDepot | 960 tubes/caps | 295.2 EUR |
Test Tube rack for 40 x 1.5/2.0 ml tubes |
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  B2000-4-T1520 | Benchmark Scientific | 1 PC | 164.65 EUR |
Test Tube rack for 40 x 1.5/2.0 ml tubes |
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B2000-4-T1520 | Benchmark Scientific | 1 each | 95.2 EUR |
Adapter 15 mL highcapacity |
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E5920716000 | Scientific Laboratory Supplies | EACH | 281.58 EUR |
Adapter 1.6 - 5 ml tubes |
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CEN0189 | Scientific Laboratory Supplies | PK6 | 71.82 EUR |
Antigen Retrieval Solution, Citrate pH 6.0 (50 ml, 100X) |
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ARS6-50 | Alpha Diagnostics | 5 L | 343.2 EUR |
Antigen Retrieval Solution, EDTA pH 8.0 (50 ml, 100X) |
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ARS8-50 | Alpha Diagnostics | 5 L | 343.2 EUR |
AXYPREP MAG DYECLEAN KIT- 50 ML - 5000 PREPS |
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MAG-DYECL-50 | CORNING | 1/pk | 1281.6 EUR |
RESERVOIR, POLYPROPYLENE, 50 ML, V-BOTTOM, NONSTERILE, BULK |
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RES-V-50 | CORNING | 100/pk | 80.4 EUR |
Adapter for 7 x 15 ml tubes conical (Falcon type) |
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CEN5048 | Scientific Laboratory Supplies | EACH | 115.14 EUR |
0.2 ml 8-Strip Low-Profile Tubes & Flat Caps, 120 Strips of 8 Tubes and Caps |
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GP201 | GenDepot | 960 tubes/caps | 262.8 EUR |
0.2 ml 8-Strip Low-Profile Tubes & Domed Caps, 120 Strips of 8 Tubes and Caps |
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GP202 | GenDepot | 960 tubes/caps | 262.8 EUR |
0.2 ml 8-Strip Tubes & Domed Caps, Assorted Colors, 250 Strips of 8 Tubes and Caps |
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GP206 | GenDepot | 2,000 tubes/caps | Ask for price |
0.2 ml 8-Strip White Tubes & Optically Clear Caps, 120 Strips of 8 Tubes and Caps |
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GP208 | GenDepot | 960 tubes/caps | 295.2 EUR |
0.2 ml 8-Strip Low Profile Tubes, 120 Strips of 8 Tubes |
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GP303 | GenDepot | 960 tubes | 230.4 EUR |
The uptake and accumulation of [3H]noradrenaline was temperature-sensitive and inhibited by the tricyclic antidepressant desipramine. Membrane vesicles isolated from PC-12 cells represent a useful model for the investigation of the molecular mechanism of the neuronal noradrenaline transport system.