DOI: 10.5281/zenodo.11395806
title: SEC-SAXS and SAXS data of human haemoglobin in 100mM NaP buffer pH=7.0 without and with PEGs.

description: The SEC-SAXS folder contains raw SEC-SAXS data of human haemoglobin in 100 mM NaP buffer, pH = 7.0.

The Hb_PEG folder contains raw SAXS data of human haemoglobin in 100 mM NaP buffer, pH = 7.0 without and with PEGs.

 
description: The SEC-SAXS folder contains raw SEC-SAXS data of human haemoglobin in 100 mM NaP buffer, pH = 7.0.

The main parameters of the instrument and measurements are presented in the file 220524Hemoglobin.Protocol.html.

uv.csv contains UV-Vis spectra of the sample across the entire SEC analysis (data from UV-Vis spectrometer in the SAXSpoint 2.0).

by_min.csv and by_ml.csv contain run-time SEC parameters (from AktaGo system).

 

Laboratory small-angle X-ray scattering (SAXS) experiments were carried out using a SAXSpoint 2.0 system from Anton Paar, Austria. This instrument features a high-intensity MetalJet C2+ X-ray source from Excillum, Sweden, that produces X-rays with a wavelength of 1.34 Å. The size of the X-ray beam incident on the sample was specified as 982.7 μm^2, and the beam had an intensity, or flux, of approximately 10^8 photons per second. X-ray images were acquired with an Eiger 1M detector from Dectris, Switzerland. The distance between the sample and the detector was 792.6 mm in SEC-SAXS measurements. The system was also equipped with an automatic sample delivery mechanism which included UV-Vis stage, CaryUV 60 from Agilent and a liquid chromatography system, the Akta Go from GE Healthcare. The acquisition time for each SAXS frame in SEC-SAXS experiment was 15 seconds, and during this time, UV-Vis spectra were recorded simultaneously to monitor the sample. 

 

The loading concentration of the sample was 18 mg/ml (approximate weight estimate) in 500 μl injection volume. The flow rate was 0.05 ml/min.

 

Abbreviations:Hb - haemoglobinNaP - sodium phosphate buffer
description: The Hb_PEG folder contains raw SAXS data of human haemoglobin in 100 mM NaP buffer, pH = 7.0 without and with PEGs.

 

The "frame_average" folder contains the average 2D files for each sample.

The .dat files are the average of 30 frames, reduced to 1D. (without buffer subtraction)

 

Laboratory small-angle X-ray scattering (SAXS) experiments were carried out using a SAXSpoint 2.0 system from Anton Paar, Austria. This instrument features a high-intensity MetalJet C2+ X-ray source from Excillum, Sweden, that produces X-rays with a wavelength of 1.34 Å. The size of the X-ray beam incident on the sample was specified as 982.7 μm^2, and the beam had an intensity, or flux, of approximately 10^8 photons per second. X-ray images were acquired with an Eiger 1M detector from Dectris, Switzerland. The distance between the sample and the detector was 825.6 mm in usual SAXS measurements. The measurements were conducted utilizing a 1 mm quartz capillary. To ensure consistency during the experiments, the samples were thermally stabilized at a temperature of 293 K (i.e., approximately 20 oC). For every sample, a total of 30 frames were collected, with each frame having an acquisition time of 30 seconds.

 

The final protein concentration in the sample is 5 mg/ml.

PEGs concentration is expressed in % (w/v).

The molecular weight of used PEGs and their concentration in the sample are noted in the names of the files.

 

Abbreviations:

Hb - haemoglobin

NaP - sodium phosphate buffer