設備名称: PETRA III EMBL P12 / 地域: Hamburg / 国: Germany / 線源: X-ray synchrotron / 波長: 0.123982 Å / スペクトロメータ・検出器間距離: 3 mm
検出器
名称: Pilatus 6M
スキャン
タイトル: Bovine serum albumin, purified monomer - SEC-SAXS coupled to multiangle laser and quasi-elastic light scattering (MALLS and QELS) 測定日: 2019年4月5日 / 保管温度: 20 °C / セル温度: 20 °C / 照射時間: 1 sec. / フレーム数: 81 / 単位: 1/nm /
Min
Max
Q
0.0979
7.213
距離分布関数 P(R)
ソフトウェア P(R): GNOM 5.0 / ポイント数: 1413 /
Min
Max
Q
0.097859
4.00096
P(R) point
1
1413
R
0
8
結果
カーブのタイプ: sec コメント: BSA monomer underwent pre-purification prior to SEC-SAXS using the following method. All procedures were performed at 4 oC. Protein powder (Sigma) consisting of BSA monomers, dimers, ...コメント: BSA monomer underwent pre-purification prior to SEC-SAXS using the following method. All procedures were performed at 4 oC. Protein powder (Sigma) consisting of BSA monomers, dimers, trimers and higher MW species was made to approximately 25 mg/ml in 25 mM HEPES, 50 mM NaCl, 5 mM urea, 1% v/v glycerol, pH 7. Approximately 200 μl of sample were loaded onto a Superdex 200 Increase 10/300 column (GE Healthcare) equilibrated in the same buffer (flow rate = 0.4 ml/min). Fractionated aliquots corresponding to the highest absorbing peak (estimated using UV A280 and UV A245 nm) were pooled and concentrated (30 kDa centrifuge spin filter) to a final concentration of 8.8 mg/ml (the concentration was determined from triplicate UV A280 measurements using an E0.1% of 0.646 (= 1 g/l) calculated from the amino acid sequence (ProtParam)). Approximately 75 μl aliquots were snap-frozen in liquid nitrogen then stored at -80oC prior to the SEC-SAXS analysis that was performed at room temperature in 50 mM HEPES, 150 mM NaCl, 2% v/v glycerol, pH 7. The Rg-correlation through the SEC-SAXS peak, the individual unsubtracted SEC-SAXS frames as well as the results from coupled MALLS and QELS analysis are included in the full entry zip archive. The quoted experimental molecular weight was determined using MALLS in combination with refractive-index (RI) measurements that were recorded from the same sample eluting from the column using a split-flow SEC-SAXS-light scattering configuration (Graewert et al., (2015) Sci. Reports. 5, 10734: doi: 10.1038/srep10734). The average hydrodynamic radius of the protein is 3.5 nm.