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

SASDBJ3 – Bovine serum albumin, monomer from SEC-SAXS

Bovine serum albumin, monomer
MWexperimental 67 kDa
MWexpected 66 kDa
VPorod 100 nm3
log I(s) 5.55×102 5.55×101 5.55×100 5.55×10-1
Bovine serum albumin, monomer small angle scattering data  s, nm-1
ln I(s)
Bovine serum albumin, monomer Guinier plot ln 5.55×102 Rg: 2.8 nm 0 (2.8 nm)-2 s2
Bovine serum albumin, monomer Kratky plot 1.104 0 3 sRg
Bovine serum albumin, monomer pair distance distribution function Rg: 2.8 nm 0 Dmax: 8.2 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Bovine serum albumin, monomer PDB (PROTEIN DATA BANK) model

log I(s)
 s, nm-1
Bovine serum albumin, monomer 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 1411-1437, corresponding to monomeric 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, monomer (BSA monomer)
Mol. type   Protein
Organism   Bos taurus
Olig. state   Monomer
Mon. MW   66.5 kDa
UniProt   P02769 (25-607)
Sequence   FASTA