Staining of Proteins & Nucleic Acid

Protein

the basic component of living cells and is made up of carbon, hydrogen, oxygen, nitrogen and 1 or more chains of amino acids linked by peptide bonds.

Simple Proteins

on hydrolysis they yield only the amino acids and occasional small carbohydrate compounds i.e. albumins, globulins, histones, protamines

Conjugated Proteins

are simple proteins combined with some non-protein material in the body to form complex proteins

i.e. lipoproteins, mucoproteins

Derived Proteins

are those derived from simple and conjugated proteins by physical or chemical means. i.e. denature proteins and peptide

Alkaline Fast Green

For basic proteins especially protamines and histones

Peracetic Acid-Alcian Blue

For cystine and cysteine

Sakaguchi’s Test

Uses NaOH, sodium hypochlorite, pyridine chloroform – this produces orange red color on objects with arginine

Sakaguchi’s Test

For arginine

Feulgen’s

Used to identify chromosomal material or DNA in cell specimens

Feulgen’s

Considered specific for DNA

Methyl Green Pyronin Method

Also utilized to detect presence of plasma cells and lymphocytes

Methyl Green Pyronin Method

Produce a differential staining reaction for DNA and RNA. Methyl green is highly selective for DNA coloring it green. Pyronin is somewhat specific for RNA giving it pinkish red color

Fluorescent Staining

May be carried out for DNA & RNA

Fluorescein

most widely used (with apple green emission)

Rhodamine

exhibits orange-red emission

Acridine orange

most commonly used fluorochrome to demonstrate DNA and RNA in fresh or fixed tissues

Acriflavine

can be used as an alternative to basic fuchsin in schiff’s reagent for the Feulgen technique