Protein's Purification & Detection methods in brief

Protein's Methods in BRIEF;

1, Protein

 1.1, Protein purification

   1.1.1, Protein isolation (Chromatography methods: ion exchange, size-exclusion chromatography (or gel filtration), affinity chromatography)

   1.1.2, Protein extraction and solubilization

   1.1.3, Concentrating protein solutions

   1.1.4, Gel electrophoresis

     1.1.4.1, Gel electrophoresis under denaturing conditions

     1.1.4.2, Gel electrophoresis under non-denaturing conditions

     1.1.4.3, 2D gel electrophoresis

   1.1.5, Electrofocusing

 1.2, Detecting proteins

   1.2.1, Non-specific methods that detect total protein only

     1.2.1.1, Absorbance (Read at 280 or 205 nm. Can be very inaccurate. Detection in the range of 100 μg/mL to 1 mg/mL. Ratio of absorbance readings taken at 260/280 can indicate purity/contamination of the sample (pure samples have a ratio <0.8))

     1.2.1.2, Bradford protein assay (Detection in the range of ~1 mg/mL)

     1.2.1.3, Biuret Test Derived Assays

       1.2.1.3.1, Bicinchoninic acid assay (BCA assay) (Detection down to 0.5 μg/mL)

       1.2.1.3.2, Lowry Protein assay (Detection in the range of 0.01–1.0 mg/mL)

     1.2.1.4, Fluorescamine (Quantifies proteins and peptides in solution if primary amine are present in the amino acids)

     1.2.1.5, Amido black (Detection in the range of 1-12 μg/mL)

     1.2.1.6, Colloidal gold (Detection in the range of 20 - 640 ng/mL)

     1.2.1.7, Nitrogen detection

       1.2.1.7.1, Kjeldahl method (used primarily for food and requires oxidation of material)

       1.2.1.7.2, Dumas method (used primarily for food and requires combustion of material)

   1.2.2, Specific methods which can detect amount of a single protein

     1.2.2.1, Spectrometry methods

       1.2.2.1.1, High-performance liquid chromatography (HPLC) (Chromatography method to detect proteins or peptides)

       1.2.2.1.2, Liquid chromatography–mass spectrometry (LC/MS) (Can detect proteins at low concentrations (ng/mL to pg/mL) in blood and body fluids, such as for Pharmacokinetics)

     1.2.2.2, Antibody dependent methods

       1.2.2.2.1, Enzyme-linked immunosorbent assay (ELISA) (Specifically can detect protein down to pg/mL)

       1.2.2.2.2, Protein immunoprecipitation (technique of precipitating a protein antigen out of solution using an antibody that specifically binds to that particular protein)

       1.2.2.2.3, Immunoelectrophoresis (separation and characterization of proteins based on electrophoresis and reaction with antibodies)

       1.2.2.2.4, Western blot (couples gel electrophoresis and incubation with antibodies to detect specific proteins in a sample of tissue homogenate or extract (a type of Immunoelectrophoresis technique))

       1.2.2.2.5, Protein immunostaining

   1.2.3, Protein structures

     1.2.3.1, X-ray crystallography

     1.2.3.2, Protein NMR

     1.2.3.3, Cryo-electron microscopy

     1.2.3.4, Small-angle X-ray scattering

     1.2.3.5, Circular Dichroism

   1.2.4, Interactions involving proteins

     1.2.4.1, Protein footprinting

     1.2.4.2, Protein–protein interactions

       1.2.4.2.1, (Yeast) two-hybrid system

       1.2.4.2.2, Protein-fragment complementation assay

       1.2.4.2.3, Co-immunoprecipitation

       1.2.4.2.4, Affinity purification

       1.2.4.2.5, mass spectrometry

       1.2.4.2.6, Proximity ligation assay

     1.2.4.3, Protein–DNA interactions

       1.2.4.3.1, ChIP-on-chip

       1.2.4.3.2, Chip-sequencing

       1.2.4.3.3, DamID

       1.2.4.3.4, Microscale thermophoresis

     1.2.4.4, Protein–RNA interactions

       1.2.4.4.1, Toeprinting assay

       1.2.4.4.2, TCP-seq

   1.2.5, Computational methods

     1.2.5.1, Molecular dynamics

     1.2.5.2, Protein structure prediction

     1.2.5.3, Protein sequence alignment (sequence comparison, including BLAST)

     1.2.5.4, Protein structural alignment

     1.2.5.5, Protein ontology (see gene ontology)

   1.2.6, Other methods

     1.2.6.1, Hydrogen–deuterium exchange

     1.2.6.2, Mass spectrometry

     1.2.6.3, Protein sequencing

     1.2.6.4, Protein synthesis

     1.2.6.5, Proteomics

     1.2.6.6, Peptide mass fingerprinting

     1.2.6.7, Ligand binding assay

     1.2.6.8, Eastern blotting

     1.2.6.9, Metabolic labeling

       1.2.6.9.1, Heavy isotope labeling

       1.2.6.9.2, Radioactive isotope labeling

source: Open access sources