Preparing monodisperse macromolecular samples for successful biological small-angle X-ray and neutron-scattering experiments.

Jeffries CM, Graewert MA, Blanchet CE, Langley DB, Whitten AE, Svergun DI, Nat Protoc 11(11):2122-2153 (2016) Europe PMC

SASDBK3 – Bovine serum albumin, dimer from SEC-SAXS

Bovine serum albumin, dimer
MWexperimental 134 kDa
MWexpected 133 kDa
VPorod 202 nm3
log I(s) 2.17×102 2.17×101 2.17×100 2.17×10-1
Bovine serum albumin, dimer small angle scattering data  s, nm-1
ln I(s)
Bovine serum albumin, dimer Guinier plot ln 2.18×102 Rg: 3.9 nm 0 (3.9 nm)-2 s2
(sRg)2I(s)/I(0)
Bovine serum albumin, dimer Kratky plot 1.104 0 3 sRg
p(r)
Bovine serum albumin, dimer pair distance distribution function Rg: 4.0 nm 0 Dmax: 12.7 nm

Data validation


Fits and models


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

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

X-ray synchrotron radiation scattering data, I(s) vs s (s = 4π sin θ/λ, where 2θ is the scattering angle) were collected from a sample of bovine serum albumin using continuous-flow size-exclusion chromatography SAXS (SEC-SAXS) in combination with RALLS/RI/UV measurements at the EMBL-P12 bioSAXS beam line on the Petra-III storage ring (Hamburg, Germany). The BSA sample (60 microlitres at 11.4 mg/ml) in 25 mM Tris 150 mM NaCl 3% v/v glycerol, pH 7.5 was injected onto a GE Healthcare S200 Increase 10/300 (24 ml) column at a flow rate of 0.4 ml/min. A total of 3600 x 1 second SAXS data frames were recorded throughout the BSA elution. After background solvent corrections, SEC-SAXS data frames 1133-1163, corresponding to dimeric BSA, were scaled and averaged to produce the data displayed in this entry. Additional files, including plots of I(0) and Rg vs frame number (determined by AUTORG) as well as the processed RALLS/RI molecular weight correlations are included in the additional files of the full entry zip archive.

Bovine serum albumin, dimer (BSA dimer)
Mol. type   Protein
Organism   Bos taurus
Olig. state   Dimer
Mon. MW   66.5 kDa
 
UniProt   P02769 (25-607)
Sequence   FASTA
 
PDB code   4F5S