Antibodies, A Laboratory Manual, Second Edition, Edited by Edward A. Greenfield



Table of Contents

Expand All | Contract All

Selecting Myeloma Cells for HGPRT Mutants with 8-Azaguanine

(Protocol summary only for purposes of this preview site)

For drug-selective media to work for hybridoma selection, myeloma cells expressing a mutation abrogating the function of their HGPRT gene (and subsequently unable to produce purines for DNA biosynthesis) are used. 8-Azaguanine is an analog of guanine that is a known inhibitor of purine nucleotide biosynthesis. It is incorporated into DNA by the HGPRTase enzyme, leading to a blockage of purine nucleotide synthesis, thereby inhibiting DNA biosynthesis (Kornbery and Baker 1992). Cells that are deficient in HGPRTase cannot incorporate 8-azaguanine in vivo and thus continue to grow. Myeloma cells should be checked periodically to ensure that they maintain drug sensitivity (see Protocol 18) and reselected for using this protocol if necessary.

Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition

Search for information about other protocols included in the book: