PROTEIN CHARACTERIZATION
Gel Permeation Chromatography (GPC) is a chromatographic technique used to determine the molecular weight distributions of large molecules, such as polymers and biomolecules. GPC is also known as Size Exclusion Chromatography (SEC).
Working Principle of GPC
Size-Based Separation:
The columns used in GPC are filled with a special porous packing material. Molecules penetrate these pores based on their size:
Large Molecules: Cannot enter the pores and elute faster.
Small Molecules: Enter the pores and remain in the column longer.
Separation Mechanism:
The movement of molecules within the column depends entirely on their physical size and is not influenced by chemical properties (e.g., polarity, charge).
Detectors:
Refractive Index Detector (RI): Commonly used to measure the concentration of the sample in solution.
UV Detector: Analyzes components with UV absorption properties.
Light Scattering Detector: Used for determining molecular weight.
Viscometer Detector: Measures the intrinsic viscosity of polymers.
Parameters Measured by GPC
Number Average Molecular Weight (Mn): The average size of molecules.
Weight Average Molecular Weight (Mw): Considers the weight contribution of larger molecules.
Polydispersity Index (PDI): Indicates the breadth of molecular weight distribution.
Specifications: 
Services
- Total protein extraction from cell lines and tissues
- Total protein quantification using the Bradford method
- SDS-PAGE protein electrophoresis
- Protein transfer to membranes
- Gel/membrane imaging
- Intensity calculation and result analysis
Flow Cytometry
Flow cytometry is a technique used to analyze the physical and chemical properties of cells and particles within a fluidic system. This method enables single-cell or particle-based analysis and is widely applied in various fields.
Features Measured by Flow Cytometry
- Cell Size and Internal Granular Structure: Determined by FSC (forward scatter) and SSC (side scatter).
- Surface and Intracellular Proteins: Measured using fluorescent dyes conjugated to antibodies.
- DNA and RNA Content: For cell cycle analysis and apoptosis studies.
- Cytokines and Secretory Products: Using intracellular staining methods.
- Apoptosis and Necrosis: Analyzed with dyes such as Annexin V and Propidium Iodide (PI).
Applications of Flow Cytometry
- Immunology Studies: Analysis of T and B cell subsets, lymphocyte proliferation assays.
- Cancer Research: Tumor cell phenotyping, DNA ploidy, and cell cycle analysis.
- Cell Function Studies: Monitoring cellular activities like oxidative stress and calcium flux.
- Hematology: Diagnosis of leukemia and lymphoma, stem cell counting.
- Microbiology: Analysis of bacterial and fungal populations.
- Pharmaceutical Research: Drug screening and monitoring cellular effects of treatments.
- Equipment: BD Accuri C6
Isothermal Titration Calorimetry (ITC)
ITC is a highly sensitive instrument used to measure the thermodynamic parameters of molecular interactions. By detecting the heat released or absorbed during binding events, this technique provides direct insights into binding kinetics and thermodynamics.
Parameters Measured by ITC
- Binding Enthalpy (ΔH): The amount of heat released or absorbed during binding.
- Binding Stoichiometry (n): The number of ligands bound per molecule.
- Binding Constant (Ka): The equilibrium constant indicating binding strength.
- Free Energy Change (ΔG): Reflects the spontaneity of the binding process.
- Entropy Change (ΔS): Indicates changes in disorder during binding.
Applications of ITC
- Protein-Ligand Interactions: Studying ligand binding to target proteins in drug development.
- Enzyme-Coenzyme Interactions: Analyzing enzyme kinetics and cofactor binding.
- Protein-Protein Interactions: Characterizing biomolecular complexes, such as antibody-antigen binding.
- DNA and RNA Interactions: Examining the binding of proteins or small molecules to nucleic acids.
- Lipid-Membrane Interactions: Investigating lipid binding to membrane proteins or membrane stability.
Equipment: Malvern Panalytical MICROCAL PEAQ – ITC
Specifications: 
Lyophilizer
A lyophilizer is a laboratory and industrial device used for freeze-drying materials. It is commonly employed to preserve biological materials, pharmaceuticals, food products, and other samples with delicate structures for extended periods without degradation. This process, known as freeze-drying, is based on the principle of sublimation, where water transitions directly from a solid to a gaseous state.
Applications of Lyophilizers
- Pharmaceutical and Biotechnology Industries: Used to enhance the stability and shelf life of vaccines, enzymes, antibiotics, proteins, and biological products. Freeze-drying is particularly favored for biopharmaceuticals to maintain biological activity.
- Food Industry: Preserves the nutritional value, flavor, and texture of food products. Examples include coffee, fruits, and instant soups.
- Research Laboratories: Ideal for drying cell culture media, sera, plasma, and other biological materials, enabling long-term storage of sensitive samples before analysis.
- Forensic Sciences: Used for the preservation of biological specimens such as blood and tissue samples.
- Cosmetic Products: Ensures the stability of sensitive cosmetic components, especially active ingredients.
- Nanotechnology and Materials Science: Preserves nanoparticles and thin films in a dry and stable environment.
Equipment: Labogene ScanVac CoolSafe
