1. (Cell lysis) 2. (Dialysis) 3. (Chromatography) 4. (Electrophoresis)

3

(Cell lysis)

3 1. (Mechanical Homogenization) 2. (Chemical) 3. (Biological)

1. (Mechanical) 1.1 Mechanical Homogenization Pestle Mortar and Pestle Homogenizer Glass Bead Homogenizer Electrical 1.2 Thermal Disintegration Homogenizer Freezing and Thawing 1.3 High Pressure Extrusing Sonic Disintegration

1.1 Mechanical Homogenization

Mortar and Pestle

Pestle Homogenizer

Electrical Homogenizer Sonic Homogenizer

2. (Chemical) - Butanol, Saponin, Digitonin 3. (Biological) - Cellulase, Collagenase Cell lysis

. (Dialysis)

(Permeable Membrane)

Salting Out ( )

1. 2. 3.

(Chromatography)

(Separation) ation) (Identification) 2 (Stationary Phase) (Mobile Phase)

(Chromatography)

1. (Non-column Chromatography)

2. (Column Chromatography)1) Adsorption Chromatography 2) Ion-exchange Chromatography 3) Gel Filtration Chromatography 4) Affinity Chromatography

1) Paper Chromatography 2) Thin-layer Chromatography

(Nom-column Chromatography)

(Paper Chromatography) (Stationary Phase) (Mobile Phase) (Supporting medium)

---> Stationary Phase

Separation

Mixtu Components reBlue Black Red Yellow

Mobile Phase

Affinity to Stationary Phase---------------

Compon Affinity to Mobile Phase entInsoluble in Mobile Phase

--->

PC chromatogramThe final chromatogram can be compared with other known mixture chromatograms to identify sample mixes. Compounds can be identified by calculating Rf (Retention).

--->

Rf - -

- Rf - -

--->

Rf1. 2. 4. 5. 3. Rf Rf 1 Rf Rf Rf

--->

1. 2. Rf 3.

--->

1. 2. - - UV -

--->

1. 2.

(Nom-column Chromatography)

(Thin-layer Chromatography) (Stationary Phase) (Mobile Phase)

(Supporting medium) Silica gel (SiO2),

Alumina (Al2O3) ,Calcium sulfate(CaSO4)

(Column Chromatography)

- Column- (Eluent) Mobile Phase

1) Adsorption Chromatography

, , (Stationary Phase) (Mobile Phase)

2) Ion-exchange Chromatography (Stationary Phase) (Resin) (Ion exchanger) (Mobile Phase)

---> Ion-exchange Chromatography

(Ion-exchanger)

---> Ion-exchange Chromatography --->

---> Ion-exchange Chromatography --->

3) Gel filtration Chromatography

(Stationary Phase) ( Polysaccharide) Sephadex, Sepharose, Bio-gel (Mobile Phase)

4) Affinity Chromatography , - (Stationary Phase) (Mobile Phase)

5) Gas-Liquid Chromatography (Stationary Phase) (Mobile Phase)

---> Gas-Liquid Chromatography

Chromatogram

Suggest identities of some of the unlabelled peaks.

6) High Performance Liquid Chromatography (Stationary Phase) (Mobile Phase)

---> High Performance Liquid Chromatography

(Electrophoresis)

Electrophoresis

Electrophoresis

-

+

Catio n

-2 +3

-

= 0

Anio n

Electrophoresis

Factors affected the mobility of molecules 1. Molecular factors Charge Size Shape 2. Environment factors Electric field strength Supporting media (pore: sieving effect) Running buffer

Electrophoresis

1. (Power supply) 2. (Supporting medium) 3. (Buffer solution) 4.

Electrophoresis

1. (Paper Electrophoresis) 2. (Cellulose Electrophoresis) 3. (Starch Gel Electrophoresis) 4. (Polyacrylmide Gel Electrophoresis) 5. (SDS-Polyacrylmide Gel Electrophoresis)

Electrophoresis

(Paper Electrophoresis)

Electrophoresis

(Cellulose Electropho

Electrophoresis

(Starch Gel Electrophoresi

Electrophoresis

(Polyacrylmide Gel Electroph

ectrophoresis ---> Polyacrylmide Gel Electrophoresis

Polyacrylamide Gels Acrylamide polymer; very stable gel can be made at a wide variety of concentrations gradient of concentrations: large variety of pore sizes (powerful sieving effect)

Electrophoresis

5. (SDS-Polyacrylmide Gel Electrophoresis)

Electrophoresis

(Agarose Gel Electrophor

ectrophoresis ---> Agarose Gel Electrophoresis

Agarose Gel purified large MW polysaccharide (from agar) very open (large pore) gel used frequently for