Tabletop Rotors and Tubes

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Rotors 0 1 Soluble Proteins Cytochrome c Collagen Albumin Luteinizing hormone Immunoglobulin G 5 2 3 4 6 7 Yeast tRNA Ribosomal subunits Ribosomes Aldolase Catalase a 2 -Macroglobulin 10 8 9 20 40 60 80 E. coli rRNA Calf liver DNA Vesicular stomatitis virus RNA Bacteriophage T5 DNA Bacteriophage T2 & T4 DNAs Broad bean mottle Nucleic Acids Polysomes 100 200 400 Poliomyelitis Tobacco mosaic Equine encephalitis Viruses Subcellular Particles Microsomes 1 000 600 800 2000 4000 Rous sarcoma Feline leukemia Bacteriophage T2 6000 8000 Plasma membranes 10 000 Mitochondria 100 000 Figure 1-4. Sedimentation Coefficients (in Svedberg Units) for Some Common Biological Materials where v d ρ p ρ c = sedimentation velocity (dr/dt) = particle diameter = particle density = solution density µ = viscosity of liquid media g = standard acceleration of gravity At equilibrium, ρ p –ρ c is zero, and particle velocity is therefore zero. Tabletop Ultracentrifuge Rotors and Tubes 1-11
Rotors The gradient may be preformed before the run or generated during centrifugation. For gradients formed by centrifugation, the time it takes to form a gradient depends on the sedimentation and diffusion coefficients of the gradient material, the pathlength, and the rotor speed. For a given gradient material, the shorter the pathlength and the higher the rotor speed, the faster the gradient will form. In general, the time required for gradients to reach equilibrium in swinging bucket rotors will be longer than in fixed angle rotors. One way to reduce run times is to use partially filled tubes. Refer to the appropriate rotor instruction manual to determine the maximum allowable speed and solution density when using partially filled tubes. RATE ZONAL SEPARATIONS Particle separation achieved with rate zonal separation is a function of the particles’ sedimentation coefficient (density, size, and shape) and viscosity of the gradient material. Sucrose is especially useful as a gradient material for rate zonal separation because its physical characteristics are well known and it is readily available. Samples are layered on top of the gradient. Under centrifugal force, particles migrate as zones. Rate zonal separation is time dependent; if the particles are more dense than the most dense portion of the gradient, some or all of the particles will pellet unless the run is stopped at the appropriate time. A separation is sometimes a combination of rate zonal and isopycnic. Depending on particle buoyant densities and sedimentation coefficients, some particles may be separated by their differential rates of sedimentation, while others may reach their isopycnic point in the gradient. Clearing factors of swinging bucket rotors at maximum speeds and various particle densities have been calculated for 5 to 20% (wt/wt) linear sucrose gradients at 5°C. These are called k′ factor, and are given in the applicable rotor manuals. These constants can be used to estimate the time, t (in hours), required to move a zone of particles of known sedimentation coefficient and density to the bottom of a 5 to 20% gradient: k t = --- ′ s (7) 1-12 Tabletop Ultracentrifuge Rotors and Tubes
Page 1 and 2: TLR-IM-9 ROTORS AND TUBES For Beckm
Page 3 and 4: Mechanical Safety • Use only comp
Page 5 and 6: SCOPE OF THIS MANUAL This manual co
Page 7 and 8: Contents Page Cellulose Propionate
Page 9 and 10: Contents Page Tube and Accessory Ca
Page 11 and 12: Tables Page Table 1-1. Rotors Used
Page 13 and 14: Rotors Fixed Angle Rotor Swinging B
Page 15 and 16: Rotors ! CAUTION In all tabletop ul
Page 17 and 18: Rotors • Near vertical tube rotor
Page 19 and 20: Rotors Table 1-1. Rotors Used in Be
Page 21: Rotors where s is the sedimentation
Page 25 and 26: Rotors ROTOR BALANCE The mass of a
Page 27 and 28: Rotors • Cesium Chloride Gradient
Page 29 and 30: Tubes and Accessories thinwall poly
Page 31 and 32: Tubes and Accessories POLYALLOMER T
Page 33 and 34: Tubes and Accessories organic solve
Page 35 and 36: Tubes and Accessories SPACERS AND F
Page 37 and 38: Using Tubes and Accessories You can
Page 39 and 40: Using Tubes and Accessories Table 3
Page 41 and 42: Using Tubes and Accessories ➠ NOT
Page 43 and 44: Using Tubes and Accessories ! CAUTI
Page 45 and 46: Using Tubes and Accessories METHOD
Page 47 and 48: Using Tubes and Accessories Polycar
Page 49 and 50: Using Tubes and Accessories Spacer
Page 51 and 52: Using Tubes and Accessories QUICK-S
Page 53 and 54: 4 Using Rotors This section contain
Page 55 and 56: Using Rotors ROTOR PREPARATION AND
Page 57 and 58: Using Rotors ! CAUTION In all table
Page 59 and 60: Using Rotors ROTOR PREPARATION AND
Page 61 and 62: Using Rotors OPERATION For runs at
Page 63 and 64: Using Rotors VERTICAL TUBE ROTORS D
Page 65 and 66: Using Rotors Rotor Preparation and
Page 67 and 68: Using Rotors 3. Refer to the centri
Page 69 and 70: Care and Maintenance CLEANING Wash
Page 71 and 72: Care and Maintenance STERILIZATION
Page 73 and 74:
Care and Maintenance TUBE AND ACCES
Page 75 and 76:
Care and Maintenance ! CAUTION Do n
Page 77 and 78:
Care and Maintenance RETURNING A RO
Page 79 and 80:
Care and Maintenance SYMPTOM POSSIB
Page 81 and 82:
Appendix B The Use of Cesium Chlori
Page 83 and 84:
The Use of Cesium Chloride Curves 1
Page 85 and 86:
The Use of Cesium Chloride Curves T
Page 87 and 88:
Appendix C Gradient Materials This
Page 89 and 90:
Gradient Materials Table C-2. Densi
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Gradient Materials Table C-4. Densi
Page 93 and 94:
Glossary of Terms clearing factor,
Page 95 and 96:
Glossary of Terms Radel polyphenyls
Page 97 and 98:
Appendix E References Documents ref
Page 99 and 100:
References DS-640 Formation of Line
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ULTRACENTRIFUGE ROTOR WARRANTY All
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Tabletop Rotors and Tubes

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