88:
23:
32:
light-sensitive diode array or by film in the older machines. The operator can thus observe the change of sample concentration versus the axis of the rotation profile with time as a result of the applied centrifugal field. With modern instrumentation, these observations are electronically digitized and stored for further mathematical analysis.
126:
to 10 GDa can be covered. Sedimentation velocity experiments can also be used to study reversible chemical equilibria between macromolecular species, by either monitoring the number and molar mass of macromolecular complexes, by gaining information about the complex composition from multi-signal
95:
An analytical ultracentrifuge has a light source and optical detectors. To allow the light to pass through the analyte during the ultracentrifuge run, specialized cells are required which have to meet high optical standards as well as to resist the centrifugal forces. Each cell consists of a housing,
26:
Spinco Model E Analytical
Ultracentrifuge. This is an early instrument, dating to the 1950s. The operator is seated in front of the sample chamber, with his left hand touching the rotor. In operation, the chamber would be sealed behind an armored shroud and pumped down to vacuum. The shroud has
31:
In an analytical ultracentrifuge (commonly abbreviated as AUC), a sample’s sedimentation profile is monitored in real time by an optical detection system. The sample is detected via ultraviolet light absorption and/or interference optical refractive index sensitive system, monitored by
121:
Sedimentation velocity experiments render the shape and molar mass of the analytes, as well as their size-distribution. The size resolution of this method scales approximately with the square of the particle radii, and by adjusting the rotor speed of the experiment size-ranges from
96:
two windows made from optically pure quartz glass, and a centrepiece with one or two sectors and filling holes for the sector(s), closed with a screw plug in the housing. These cell are placed into a rotor cavity with a continuous bore, with a collar at the bottom to retain the cell.
112:
By applying specific equipment and adapting measurement parameters several types of experiments can be performed. Most common AUC experiments are sedimentation velocity and sedimentation equilibrium experiments.
127:
analysis exploiting differences in each components spectroscopic signal, or by following the composition dependence of the sedimentation rates of the macromolecular system, as described in
146:
constants. The rotor speed is adjusted such that a steady-state concentration profile c(r) of the sample in the cell is formed, where sedimentation and diffusion cancel out each other.
67:, which depends upon the size of the molecules being sedimented. This is the ratio of a particle's sedimentation velocity to the applied acceleration causing the sedimentation.
70:
Analytical ultracentrifugation has recently seen a rise in use because of increased ease of analysis with modern computers and the development of software, including a
39:, conformational changes in macromolecules, and size distributions of macromolecules. With AUC it is possible to gain information on the number and subunit
328:
185:
307:
319:
493:
488:
71:
87:
180:
64:
91:
Rotors for an
Analytical Ultracentrifuge for a maximal spinning velocity of 50,000 (left) and 60,000 (right) rpm
503:
300:
Reversible
Associations in Structural and Molecular Biology (RASMB -an Analytical Ultracentrifugation Forum)
128:
190:
387:"Studying multiprotein complexes by multisignal sedimentation velocity analytical ultracentrifugation"
483:
398:
143:
48:
63:
or other assemblies from different molecule classes. The simplest measurement to be obtained is the
35:
The information that can be obtained from an analytical ultracentrifuge includes the gross shape of
498:
461:
426:
373:
278:
19:
is an analytical technique which combines an ultracentrifuge with optical monitoring systems.
451:
416:
406:
363:
355:
268:
260:
175:
123:
323:
311:
165:
304:
402:
22:
456:
439:
368:
343:
273:
248:
170:
421:
386:
477:
210:
44:
40:
36:
316:
142:
Sedimentation equilibrium experiments reports the molar mass of analytes and their
134:
The experiment aims to monitor the sedimentation behavior at a fixed angular speed.
237:
C.M. Smales and D.C. James, Eds. Volume 308: 301-318. Humana Press Inc, Totowa, NJ.
60:
385:
Balbo A, Minor KH, Velikovsky CA, Mariuzza RA, Peterson CB, Schuck P (Jan 2005).
264:
440:"Analytical ultracentrifugation as a contemporary biomolecular research tool"
344:"Modern analytical ultracentrifugation in protein science: A tutorial review"
411:
305:
465:
430:
377:
282:
249:"The analysis of macromolecular interactions by sedimentation equilibrium"
359:
299:
52:
86:
21:
27:
been lowered to allow access for loading or unloading the rotor.
56:
211:"Technical Manual, Spinco Ultracentrifuge Model E"
253:Modern Analytical Ultracentrifugation: Methods
233:Perez-Ramirez, B. and Steckert, J.J. (2005).
215:Science History Institute Digital Collections
8:
235:Therapeutic Proteins: Methods and Protocols.
342:Lebowitz J, Lewis MS, Schuck P (Jan 2005).
455:
420:
410:
367:
272:
329:Report on an ultracentrifuge explosion.
202:
7:
74:supported software package, SedFit.
186:Buoyant density ultracentrifugation
14:
150:Density gradient centrifugation
17:Analytical ultracentrifugation
1:
438:Cole J, Hansen J (Dec 1999).
72:National Institutes of Health
265:10.1016/j.ymeth.2010.12.005
181:Differential centrifugation
520:
51:of macromolecules such as
138:Sedimentation equilibrium
65:sedimentation coefficient
412:10.1073/pnas.0408399102
317:Gilbert-Jenkins theory
247:Ghirlando, R. (2011).
129:Gilbert-Jenkins theory
117:Sedimentation velocity
92:
28:
494:Laboratory techniques
191:Zippe-type centrifuge
90:
49:equilibrium constants
25:
489:Analytical chemistry
144:chemical equilibrium
108:Types of experiments
403:2005PNAS..102...81B
360:10.1110/ps.0207702
322:2007-05-01 at the
310:2002-08-04 at the
93:
29:
511:
469:
459:
434:
424:
414:
381:
371:
354:(9): 2067–2069.
287:
286:
276:
244:
238:
231:
225:
224:
222:
221:
207:
176:Theodor Svedberg
519:
518:
514:
513:
512:
510:
509:
508:
474:
473:
472:
437:
384:
348:Protein Science
341:
337:
335:Further reading
324:Wayback Machine
312:Wayback Machine
296:
291:
290:
246:
245:
241:
232:
228:
219:
217:
209:
208:
204:
199:
166:Ultracentrifuge
162:
157:
155:Data evaluation
152:
140:
119:
110:
105:
99:
85:
83:Instrumentation
80:
12:
11:
5:
517:
515:
507:
506:
504:Fluid dynamics
501:
496:
491:
486:
476:
475:
471:
470:
435:
382:
338:
336:
333:
332:
331:
326:
314:
302:
295:
294:External links
292:
289:
288:
259:(1): 145–156.
239:
226:
201:
200:
198:
195:
194:
193:
188:
183:
178:
173:
171:Gas centrifuge
168:
161:
158:
156:
153:
151:
148:
139:
136:
118:
115:
109:
106:
104:
101:
84:
81:
79:
76:
47:complexes and
37:macromolecules
13:
10:
9:
6:
4:
3:
2:
516:
505:
502:
500:
497:
495:
492:
490:
487:
485:
482:
481:
479:
467:
463:
458:
453:
450:(4): 163–76.
449:
445:
444:J Biomol Tech
441:
436:
432:
428:
423:
418:
413:
408:
404:
400:
396:
392:
388:
383:
379:
375:
370:
365:
361:
357:
353:
349:
345:
340:
339:
334:
330:
327:
325:
321:
318:
315:
313:
309:
306:
303:
301:
298:
297:
293:
284:
280:
275:
270:
266:
262:
258:
254:
250:
243:
240:
236:
230:
227:
216:
212:
206:
203:
196:
192:
189:
187:
184:
182:
179:
177:
174:
172:
169:
167:
164:
163:
159:
154:
149:
147:
145:
137:
135:
132:
130:
125:
116:
114:
107:
102:
100:
97:
89:
82:
77:
75:
73:
68:
66:
62:
61:nanoparticles
58:
54:
50:
46:
42:
41:stoichiometry
38:
33:
24:
20:
18:
447:
443:
397:(1): 81–86.
394:
390:
351:
347:
256:
252:
242:
234:
229:
218:. Retrieved
214:
205:
141:
133:
120:
111:
98:
94:
69:
34:
30:
16:
15:
484:Centrifuges
499:Biophysics
478:Categories
220:2018-12-18
197:References
122:100
466:19499023
431:15613487
378:12192063
320:Archived
308:Archived
283:21167941
160:See also
53:proteins
45:covalent
457:2291609
399:Bibcode
369:2373601
274:3090454
78:History
43:of non-
464:
454:
429:
422:538923
419:
376:
366:
281:
271:
103:Theory
462:PMID
427:PMID
391:PNAS
374:PMID
279:PMID
452:PMC
417:PMC
407:doi
395:102
364:PMC
356:doi
352:102
269:PMC
261:doi
57:DNA
480::
460:.
448:10
446:.
442:.
425:.
415:.
405:.
393:.
389:.
372:.
362:.
350:.
346:.
277:.
267:.
257:58
255:.
251:.
213:.
131:.
124:Da
59:,
55:,
468:.
433:.
409::
401::
380:.
358::
285:.
263::
223:.
Text is available under the Creative Commons Attribution-ShareAlike License. Additional terms may apply.
