PISA screening protocol - A protocol for the selection of intracellular antibodies against post-translational modifications of native proteins
Protein post-translational modifications (PTMs) represent an untapped source of targets of huge biological interest and therapeutic potential. Their validation can only be indirect, because PTMs cannot be selectively hit by nucleic-acid based interference approaches. Here we describe the protocol for Post-translational Intracellular Silencing Antibody technology (P.I.S.A.), a method for the selection of intrabodies targeting the native acetylated version of proteins and their use for PTM-selective targeting and functional interference in living cells.
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Table 1 Sequences of validated acetylase domains and linkers Sequences of validated acetylase domains and linkers
Reagents, media, solutions, equipment Reagents, media, solutions, equipment
PISA screening protocol PISA screening protocol
In the Reagents section the YPA medium has the same composition of the YPAD one.
Posted 17 Jan, 2017
PISA screening protocol - A protocol for the selection of intracellular antibodies against post-translational modifications of native proteins
Posted 17 Jan, 2017
Protein post-translational modifications (PTMs) represent an untapped source of targets of huge biological interest and therapeutic potential. Their validation can only be indirect, because PTMs cannot be selectively hit by nucleic-acid based interference approaches. Here we describe the protocol for Post-translational Intracellular Silencing Antibody technology (P.I.S.A.), a method for the selection of intrabodies targeting the native acetylated version of proteins and their use for PTM-selective targeting and functional interference in living cells.
Figure 1
Figure 2
In the Reagents section the YPA medium has the same composition of the YPAD one.
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