Environmental Engineering Reference
In-Depth Information
BAM
N
-
tert
-butylacrylamide
PIC Protease inhibitor cocktail
PMSF Phenyl methyl sulfonyl fluoride
PTMs Posttranslational modifications
SPE Serum protein electrophoresis
SDS Sodium dodecyl sulphate
TCA Trichloroacetic acid
KOH Potassium hydroxide
MPS Mononuclear phagocytic system
MS Mass spectroscopy
MWCNTs Multi-walled carbon nanotubes
NIPAM
N
-isopropylacrylamide
SILAC Stable Isotope labelling by amino acids in cell culture
ALP Alkaline phosphatase
LDH Lactate dehydrogenase
CK Creatine kinase
B. garinii Borrelia garinii
C. trachomatis Chlamydia trachomatis
E. coli
Escherichia coli
H. pylori
Helicobacter pylori
15.1
Introduction
15.1.1 Electrophoresis
Electrophoresis is widely used in separation and purification of macromolecules,
e.g. proteins and nucleic acids of different size and charge. The charged molecules
migrate towards either the positive or negative pole according to their charge in
presence of electric field. The gel used in electrophoresis is composed of either
agarose (0.5-2 %) or polyacrylamide (3.5-20 % in 14
14-cm) slabs or as
minislabs (6
8-cm). Polyacrylamide gels have a rather small range of separation,
but very high resolving power than agarose gels. The separation of biomolecules by
electrophoresis mainly depends on the experimental system, properties of biomol-
ecules, namely net charge, mass and shape. Gel placement could be as slab or as
filled tube.
The protein separation can be native based on charge and mass denaturing
(particle size dependant), buffer system based and other parameters inclined tech-
niques (IEF based, 2DE). Native system can be divided into discontinuous and
continuous mode, denaturing system can be classified as SDS based and urea
formamide based to denature protein while others are based on separation based
on IEF in pH gradient. The 2DE is a combination of IEF and SDS. Generally
vertical apparatus is used in PAGE while horizontal apparatus in agarose, starch
gel, and IEF. Evaluation of gel electrophoresis over the years (Fig.
15.1
).
