BioHPLC Column Selection Guide Cover - Agilent Technologies
BioHPLC Column Selection Guide Cover - Agilent Technologies
BioHPLC Column Selection Guide Cover - Agilent Technologies
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size exclusion Chromatography (seC)<br />
Calibration of the ProSEC 300S column<br />
with globular proteins<br />
Mobile Phase: 50 mM KH 2PO 4-K 2HPO 4 (@ pH 6.8)<br />
containing 0.3 M NaCl<br />
Flow Rate: 1.0 mL/min<br />
Detector: UV, 280 nm<br />
Sample: Protein samples<br />
Molecular weights of the proteins.<br />
Mw/Daltons Protein<br />
670,000 Thyroglobulin<br />
155,000 γ-Globulin<br />
66,430 Bovine serum albumin<br />
44,287 Ovalbumin<br />
29,000 Carbonic anhydrase<br />
16,700 Myoglobulin<br />
12,384 Cytochrome c<br />
1,423 Bacitracin<br />
Analysis of Bovine Serum Albumin<br />
by light scattering using ProSEC 300S columns<br />
<strong>Column</strong>: ProSEC 300S<br />
PL1147-6501<br />
7.5 x 300 mm, 5 µm<br />
Mobile Phase: Water + 120 mM NaCl, 2.7 mM KCI, 10 mM NaH 2PO 4<br />
Flow Rate: 1.0 mL/min<br />
Detector: Differential refractive index<br />
+ 15/90 dual angle light scattering<br />
Sample: Bovine serum albumin<br />
Molecular Weights<br />
Monomer 66,900 Daltons, 88.5%<br />
Dimer 34,900 Daltons<br />
(2.02 x monomer molecular weight), 9.8%<br />
Trimer 197,000 Daltons<br />
(2.94 x monomer molecular weight), 1.2%<br />
Tetramer 279,300 Daltons<br />
(5.17 x monomer molecular weight), 0.5%<br />
Overlay of differential refractive index and dual angle light scattering sample.<br />
www.agilent.com/chem/biohplc<br />
Log M<br />
6<br />
5.5<br />
5<br />
4.5<br />
4<br />
3.5<br />
3<br />
5 7 9 10<br />
Retention time (min)<br />
Response<br />
0<br />
RI<br />
15 Deg<br />
90 Deg<br />
Tetramer<br />
Aggregation<br />
Trimer<br />
Monomer<br />
Dimer<br />
Retention time (min)<br />
13<br />
PSEC300Sglob<br />
14<br />
BovineSerAlb
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