Scribd
Upload
368 views21 pages

Protein Precipitation Methods

This document discusses protein precipitation through denaturation and other means. [1] Denaturation disrupts protein structures like hydrogen bonds and causes proteins to aggregate and precipitate out of solution. [2] Strong acids, bases, salts, solvents and heat can induce irreversible or reversible denaturation. [3] Precipitation can also be achieved through the addition of reagents like metallic salts, alkaloids, or tannic acid which bind to and precipitate proteins.

Uploaded by

Charisa Antonette Huelva
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
368 views21 pages

Protein Precipitation Methods

This document discusses protein precipitation through denaturation and other means. [1] Denaturation disrupts protein structures like hydrogen bonds and causes proteins to aggregate and precipitate out of solution. [2] Strong acids, bases, salts, solvents and heat can induce irreversible or reversible denaturation. [3] Precipitation can also be achieved through the addition of reagents like metallic salts, alkaloids, or tannic acid which bind to and precipitate proteins.

Uploaded by

Charisa Antonette Huelva
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd

Precipitation of Protein

Done by : Sahar Al-Subaie


Introduction
Denaturation definition:

 is a process in which proteins or nucleic acids lose


the quaternary structure, tertiary
structure and secondary structure which is present
in their native state, by application of some external
stress or compound such as a strong acid or base, a
concentrated inorganic salt, an organic solvent
(e.g., alcohol or chloroform), radiation or heat.
Denaturation:

When the structures of native proteins are altered


by chemical or physical means, the protein
molecules tend to agglomerate and precipitate
.and the protein becomes denaturated
Denaturation:
Protein precipitation is due to a
disruption of hydrogen bonds, ionic bonds
and sometimes the stronger covalent
disulfide bonds
Denaturation types

irreversible (excessive heating or


extreme PH changes)
reversible (treatment with
ammonium sulfate).
Influence of Strong Mineral
Acids:

Proteins are amphiprotic substance


.(can accept or give protons)
Influence of Strong Mineral
Acids:
The protein will precipitate because
the ionic bonds and hydrogen bonds
are disrupted.
The procedure:

Test Tube No. 1 2 3


1% egg albumin 3 ml 3 ml 3 ml
Con. HCL 2 ml x x
Con. HNO3 x 2 ml x
Con. H2SO4 x x 2 ml
Precipitation by Alkaloidal
Reagent
Alkaloidal reagents are acids that can
combine with alkaloids (Alkaloids are
.organic bases from plants)
Precipitation by Alkaloidal
Reagent

Certain acidic reagents (alkaloidal reagents) e.g :


Trichloroacetic acid, Tannic acid, Phosphotungstic acid,
Picric acid, Sulfosalicylic acid combine with protein to
form insoluble protein salts (e.g Protein tannate).
The procedure:

Test Tube No. 1 2 3 4 5 6


1%egg albumin 3 ml 3 ml 3 ml 3 ml x x
1% urea x x x x 3 ml x
1%alanine x x x x x 3 ml
10%Trichloroacetic 5 drops x x x 5 drops 5 drops
acid
5%aqueous tannic x 5 drops x x x x
acid
20%phosphotungstic x x 5 drops x x x
acid
20%sulfosalicylic x x x 5 drops x x
acid
Precipitation by Metallic Salts
When heavy metal cations (e.g: Pb, Cu, Hg, Ag)
are added to protein solution, the metal ions
combine with the negatively charged groups to
form insoluble metal ion proteinate.
The procedure:

Test Tube No. 1 2 3 4


1%egg 2 ml 2 ml 2 ml x
albumin
1%urea x x x 2 ml
0.2M copper 5 drops x x 5 drops
sulfate
0.2M lead x 5 drops x x
acetate
0.2M mercuric x x 5 drops x
chloride

You might also like