Beer–Lambert law might not be the case any more in samples with high absorbance.

TLS Online TPP Program

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#Unit 3. Fundamental Processes

Eukaryotic nuclei contain RNA polymerase I, RNA polymerase II, and RNA polymerase III can be separated by using DEAE-Sephadex ion-exchange chromatography

TLS Online TPP Program

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#Unit 3. Fundamental Processes

RNA Polymerase can also separated by using epitope tagging. Normally RPB3 is used to create epitope tag.

TLS Online TPP Program

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#Unit 3. Fundamental Processes

Molecular genetic techniques are used to make a cDNA encoding a recombinant protein to which is fused a short sequence of amino acids called an epitope tagging.

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#Unit 3. Fundamental Processes

Two commonly used epitope tags are FLAG, which encodes the amino acid sequence DYKDDDDK (single-letter amino acid code), and myc, which encodes the sequence EQKLISEEDL.

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#Unit 3. Fundamental Processes

TLS Online TPP Program

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#Unit 3. Fundamental Processes

Fluorochrome-coupled monoclonal antibodies to the FLAG or myc epitopes can then be used to detect the recombinant protein in the cell.

Alexa- 488-labeled antibody -emit green light

Alexa-568-labeled antibody- emit red light