Sub-cellular Components Flashcards
the isolation of organelles by the use differential centrifugation
Subcellular Fractionation
Steps in Subcellular Fractionation
- Extraction
- Homogenization
- Centrifugation
isolation of subcellular components by dropping specimen to sucrose or NSS
Extraction
Preferred Concentration for Sucrose Sol’n
0.025 M
Preferred Concentration for NSS
0.16 M
Reason for using Sucrose and NSS to preserve sample
Because the molecules does not cross the cells
the process of releasing the organelles inside the cells
Homogenization
Types of Homogenization
Sonication
Detergent
French Press
Mechanical Homogenization
a type of homogenization that uses sound waves
Sonication
a type of homogenization that promotes the lysis of the cell to release the oragnelles
Detergent
homogenization using pressure
French Press
the use of blender to release the organelles
Mechanical Homogenization
the process of separating the components by the use of centrifugal force at repeated and progressive speeds
Centrifugation
The smaller the COMPONENT?
The higher centrifugal force required to sediment it
Types of Supernatant
I - Nuclear Fraction
II - Mitochondrial Fraction
III - Microsomal Fraction
I - Nuclear Fraction
- 10 mins
- Nucleic Acids
- DNA, RNA
II - Mitochondrial Fraction
- 20 mins
- Mitochondria (CARBS)
- Lysosome and Golgi bodies
III - Microsomal Fraction
- more than 40 mins
- microsome (LIPIDS AND PROTEINS)
- ribosome, cytoplasm, endoplasmic reticulum
Different Qualitative Analysis for CARBS
BIURET SUDAN IV MOLISCH DISCHE BIAL/ORCINOL
detects peptide bond by removing the nitrogen of the bonds and bonding it to the Cu2+ in the reagent
Biuret’s Test
hydrophobic dye that stains lipids giving it a red color and sinks in polar substances or with the absence of lipids
Sudan IV
general test for carbs that uses a-napthol and conc. H2SO4 to form a purple ring in the sample
Molisch Test
used for the detection of DNA by using diphenlyamine to react with the 2-deoxypentose of the DNA
Dische Test
used to detect pentose such as ribose by using 3,5-dihydroxytoluene creating a blue-green appearance
Orcinol/ Bial’s Test
