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process involving multiple centrifugation steps. It increases the speed of centrifugation each time to separate cellular materials.
Centrifuge is a machine that uses centrifugal force to separate substances or parts of substances
+ result: purple ring at the junction
+ result: rose-pink to violet, then purple solution
+ result: blue solution
+ result: red solution
Glycosidase: an enzyme that catalyzes the hydrolysis of glycosidic linkages in sugar molecules.
Protease: an enzyme that catalysis the hydrolysis of peptide bonds.
Acid phosphatase: a phosphatase, an enzyme that catalyzes they hydrolysis of phosphate linkages in a variety of molecules.
Sulfatase: an enzyme that catalyzes the hydrolysis of sulfate ester linkages in molecules.
Lipase: an enzyme that catalyzes the hydrolysis of ester bonds in lipid molecules.
Amylase: an enzyme that catalyzes the breakdown of carbohydrates into sugars.
Nuclease: an enzyme that catalyzes the hydrolysis of phosphodiester bonds in nucleotide subunits of nucleic acids.
glucocerebrosidase: an enzyme which breaks down glucocerebroside
Phosphoric Acid monoesters
The energy currency of the cell (ATP or Adenosine triphosphate) is made in the mitochondria during aerobic respiration and so "powers" the cell's activities.
Sedimentation depends on mass, shape, and partial specific volume of a macromolecule, as well as solvent density, rotor size and rate of rotation.
1 ml of NaOH
+
1 ml of copper sulfate
1 ml of conc. Sulfuric acid
+
1 ml of Molisch Reagent
Molisch Reagent: α-Naphthol dissolved in Ethanol
I. Preparation of supernatant and sediment of the subcellular components of chicken liver
add Small amount of sudan black crystals
RNA
1 ml of
orcinol
reagent
+
20 min
water bath
DNA
1 ml of
Diphenylamine
reagent
+
1 ml of conc
sulfuric acid
+
20 min water
bath
II. Qualitative tests
a) Proteins (Biuret Test)
b) Lipids
c) Carbohydrates (Molisch Test)
d) Nucleic Acids (DNA)
e) RNA (Orcinol Test)
OBJECTIVES:
At the end of the experiment, the student should be able to:
Understand and appreciate the different biochemical systems that bring about, activate and maintain life
Determine the different biomolecules
present in the cell
Give a concise account of the components of cells and how they may be isolated