The Service of Protein Chemistry at the Margarita Salas Center for Biological Research (CIBMS) offers scientific and technical support in the protein investigaton to any research laboratory from public research organisms or private companies. In the Service it can be performed separation, quantification, identification and characterization of proteins by chromatography techniques, amino acid analysis, amino terminal sequencing, LC-MS and isothermal titration calorimetry. Additionally, the facility provides peptide synthesis and purification.
The service is a part of the laboratory network of the Community of Madrid. Since 2011 the laboratory is certified according to UNE-EN ISO9001:2015 with ER-0286/2009 AENOR certification code in "Automated amino acid analysis using post column ninhydrin derivatization" and "Automated protein sequencing by Edman degradation chemistry" field of activities.
The Service has the following equipment:
Protein sequencer (Applied Biosystems, Procise 494). The Procise 494 Protein Sequencer performs automated Edman sequencing chemistry. Samples can be analyzed either in solution or blotted to PVDF after PAGE separation. Sensitivity is in the low pmol range.
Amino acid analyzer (Biochrom 30). The Biochrom 30 uses the classical amino acid analysis methodology based on ion-exchange liquid chromatography and postcolumn continuous reaction with ninhydrin to provide qualitative and quantitative compositional analysis. Sensitivity is ∼10 pmol.
Peptide synthesizer (AAPPtec, Focus XC). The Focus XC can synthesize up to six peptides simultaneously at scales from 0.05-5 mmol, using Fmoc chemistry.
HPLC: ÄKTA basic (Amersham Pharmacia Biotech) and Agillent 1200 preparative purification platform.
MicroCal PEAK-ITC isothermal titration calorimeter (Malvern Panalytical). This technique enables cuantitative determination of binding constants, reaction stoichiometry, enthalpy and entropy when two molecules interact.
LC-MS. Mass spectrometry system (Thermo Mod. FinniganTM LXQ TM) with on line HPLC Surveyor with the following configuration:
- Pump with four separate solvent feeds (Surveyor MS pump Plus).
- Autosampler for multi sample analysis (Surveyor AS Plus).
- Photo Diode Array detector (Surveyor PDA Plus).
- Mass spectrometer LXQ equipped with linear ion trap. Mass range from 50-4000 m/z. Ionization sources: electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI).
Users interested in using the Scientific Service of Protein Chemistry can contact in the email address firstname.lastname@example.org or by phone 918373112, extension 4294.
All samples must be accompanied by a fully completed sample submission form, otherwise samples will not be processed. The forms are available on this website and can be sent electronically or preferably printed, signed and enclosed along with the sample.
Before shipping the samples, send us an email or call us to alert us for the arrival of the samples.
Please, send your samples in a padded envelope or in a box together with the submission form. The samples should be in an Eppendorf reaction tube sealed by Parafilm. Samples with biological or chemical risk are not accepted.
The facility is not responsible for the results obtained if the samples don´t have the suitable conditions specified in the “Sample preparation procedure” section.
Solicitud síntesis de péptidos
N-terminal sequencing request form
Solicitud análisis de aminoácidos
Relación de muestras análisis de aminoácidos
Intro Sample preparation Helpful Edman Degradation sample preparation protocols Limiting factors Applications
AMINO ACIDS ANALYSIS
Intro Sample preparation Hydrolysis method and limiting factors Helpful hints to reduce amino acid sample contamination Applications
Sample preparation Analysis conditions Forbidden additives Tolerable additives Applications
ISOTHERMAL TITRATION CALORIMETRY (ITC)