In the Isoelectric Focusing pH gradient is formed by the introduction into the gel of compounds known as ampholytes, which are complex mixtures of synthetic polyamino-polycarboxylic acids Ampholytes can be purchased in different pH ranges covering a wide band (e.g. pH 3–10).

TLS Online TPP Program

#Id: 2860

#Unit 13. Methods in Biology

Carotenoids gives absorbance between 400 – 500 nm of wavelength while porphyrin gives absorbance at 400 nm of wavelength known as Soret band.

TLS Online TPP Program

#Id: 2861

#Unit 13. Methods in Biology

DNA gives absorbance at 595 nm in presence of Diphenylamine while gives direct absorbance at 260 nm

TLS Online TPP Program

#Id: 2862

#Unit 13. Methods in Biology

RNA gives absorbance at 595 nm in presence of Diphenylamine while gives direct absorbance at 540 nm.

TLS Online TPP Program

#Id: 2863

#Unit 13. Methods in Biology

The electronic transitions of the peptide bond occur in the far UV. The intense peak at 190 nm, and the weaker one at 210–220 nm is due to the p ® p* and n ® p* transitions.

TLS Online TPP Program

#Id: 2864

#Unit 13. Methods in Biology

A number of amino acids (Asp, Glu, Asn, Gln, Arg and His) have weak electronic transitions at around 210 nm Usually, these cannot be observed in proteins because they are masked by the more intense peptide bond absorption.

TLS Online TPP Program

#Id: 2865

#Unit 13. Methods in Biology

The most useful range for proteins is above 230 nm, where there are absorptions from aromatic side chains.