Proteins

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Proteins by Mind Map: Proteins

1. Maintain metabolic processes in cell

2. Convey information about extracellular environment of cell

2.1. Mediate effect of receptors : INTRACELLULAR SIGNALLING COMPONENT

3. catalysts

4. receptors

5. DNA&RNA

5.1. DNA REPLICATION, DNA RECOMBINATION, RNA SPLICING OR EDITING

6. sample preparation

6.1. cell disruption

6.1.1. To release proteins from within cell

6.1.1.1. Mechanical method

6.1.1.2. Non-mechanical method

6.2. protein solubilization

6.2.1. To denature protein

6.2.2. To disrupt disulfide bond through reduction

6.2.3. To dissolve particles interfere with electrophoresis

6.2.3.1. Sample/Loading buffer

6.2.3.1.1. Tris-Cl

6.2.3.1.2. Glycerol

6.2.3.1.3. Sodium Dodecyl Sulfate (SDS)

6.2.3.1.4. B-mercaptoethanol

6.2.3.1.5. Bromophenol blue

6.3. contaminant removal

6.3.1. Remove interfering substances that can negatively impact SDS-PAGE

6.3.1.1. Buffer Exchange (Desalting)

6.3.1.2. Protein Precipitation ( Help concentrate sample if needed)

6.4. quantitation

6.4.1. BCA protein assay : Lowry, Bradford, Biuret

6.4.1.1. Concentration by spectrofotometer

6.4.2. Related to H atom, Dalton

6.4.2.1. Molecular Weight Markers

6.4.2.2. Migration pattern : gel type, gel concentration, running buffer

6.5. PAGE preparation

6.5.1. Dilute sample in appropriate sample buffer to a final sample buffer concentration of 1X

7. Size estimation

7.1. Distance migrate in mm (x-axis) against molecular mass of prestained protein band in kD (y-axis)

7.1.1. Linear Graph