Size-exclusion chromatography small-angle X-ray scattering of water soluble proteins on a laboratory instrument.

Bucciarelli S, Midtgaard SR, Nors Pedersen M, Skou S, Arleth L, Vestergaard B, J Appl Crystallogr 51(Pt 6):1623-1632 (2018) Europe PMC

SASDED5 – Bovine serum albumin monomer from synchrotron SEC-SAXS

Bovine serum albumin
MWexperimental 63 kDa
MWexpected 66 kDa
VPorod 84 nm3
log I(s) 4.88×100 4.88×10-1 4.88×10-2 4.88×10-3
Bovine serum albumin small angle scattering data  s, nm-1
ln I(s)
Bovine serum albumin Guinier plot ln 4.89×100 Rg: 2.7 nm 0 (2.7 nm)-2 s2
(sRg)2I(s)/I(0)
Bovine serum albumin Kratky plot 1.104 0 3 sRg
p(r)
Bovine serum albumin pair distance distribution function Rg: 2.7 nm 0 Dmax: 8.7 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Bovine serum albumin PDB model

log I(s)
 s, nm-1
Bovine serum albumin DAMFILT model

Synchrotron SAXS data from solutions of Bovine serum albumin monomer using SEC-SAXS in PBS plus 1 mM DTT, pH 7.4 were collected on the BM29 beam line at the ESRF storage ring (Grenoble, France) using a Dectris Pilatus 1M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.099 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle).

The BSA sample (500 microlitres at 8.1 mg/ml) was injected onto a GE Healthcare S200 Increase 10/300 (24 ml) column at a flow rate of 0.7 ml/min at 10°C. A total of 1850 x 1 second SAXS data frames were recorded throughout the BSA elution. After background solvent corrections, SEC-SAXS data frames 1221 - 1260, corresponding to monomeric BSA, were scaled, averaged and binned logarithmically to produce the data displayed in this entry. Additional files, including plots the selected frames and of I(0), HPLC UV trace and Rg vs. time (determined by AUTORG) are included in the additional files of the full entry zip archive.

Bovine serum albumin (BSA)
Mol. type   Protein
Organism   Bos taurus
Olig. state   Monomer
Mon. MW   66.4 kDa
 
UniProt   P02769 (25-607)
Sequence   FASTA
 
PDB code   4F5S