Equipment

Services include screening of the optimization of samples of specific interest and screening. Both qualitative and quantitative analysis will be provided.

Circular Dichroism

  • buffer exchange of sample into compatible buffer and concentration of sample if needed
  • loading sample into appropriate quartz cell and collection of CD spectra. Each sample has its own quartz cell specific.
  • Recording the CD spectra with optimal parameters and temperatures (if temperature dependent studies needed).
  • saving the spectra and plotting of data and computational analysis of secondary structure
  • preparing report of results

A single, per sample charge will apply for the CD instrument.

Non-instrument CD services:

  • CD Sample Prep. This optional service will provide buffer exchange (into a sodium-free buffer) and protein concentration of user-provided samples.
  • CD data analysis. This option will provide data analysis of collected CD spectra. Data will be processed using a multi-component algorithm to estimate the fraction of secondary structural elements within a protein sample.

Electron Paramagnetic Resonance Spectroscopy

  • spin-labeling and concentration of samples as needed
  • loading samples into capillaries (quartz and TPX)
  • Optimizing the Hamiltonian parameters and temperatures (in case of LT studies).
  • Recording and collection of spectra
  • plotting of spectra and measurement of line widths and determining the spin parameters.
  • For power saturation measurements, each sample is examined under three conditions (air, N2, and NiEDDA). Collected spectral data is computationally processed for P1/2 and collisional frequency values.

The EPR instrument services are categorized under three different service types, each with a defined recharge rate:

  • Simple room temperature (RT) scan. This is the lowest rate, as scans can be obtained in a matter of minutes with little instrumentation set-up time.
  • Simple low temperature (LT) scan. This rate is higher than the RT scan due to the increased time for liquid nitrogen cooling and cryostat operation. Service also includes flash freezing.
  • EPR power saturation scanning. This service will provide scanning of samples over successive power levels under three conditions (relaxer-free, hydrophilic relaxer, hydrophobic relaxer). This rate is the highest as it is the most time consuming.

Non-instrument EPR services:

  • EPR Sample Prep and Spin-labeling. This optional protein chemistry service will provide spin-labeling and protein concentration of user-provided samples.
  • EPR data analysis. This option will provide data analysis of collected EPR spectra. For protein samples, spectra will be fit by for correlation times of mobile components. For membrane samples, line-widths will be measured to estimate correlation times and order parameters. For power saturation samples, collision frequencies will be calculated for each relaxer.