TY  - GEN
T1  - An antibody staining protocol variation for nematodes that adds heat-induced antigen retrieval (HIAR)
AU  - Loer, Curtis
AU  - Witte, Hanh
AU  - Sommer, Ralf J
AU  - Hobert, Oliver
DO  - 10.17912/micropub.biology.000135
UR  - http://beta.micropublication.org/journals/biology/micropub-biology-000135/
AB  - In the nematode satellite model organism Pristionchus pacificus, transgenics and CRISPR/Cas9 have been implemented in the last decade (Schlager et al., 2009; Cinkornpumin & Hong, 2011; Witte et al., 2015; Namai & Sugimoto, 2018). While CRISPR has been used to make mutants and to knock-in short sequences, sequence insertion to date is limited to ~150 base pairs. Therefore, following expression from native loci in P. pacificus currently depends on epitope-tagging the gene of interest via CRISPR and template-dependent repair, and then using immunocytochemistry with its maddening, nearly infinite possible variations in fixation, permeabilization, blocking, and antibody incubation protocols.
PY  - 2019
JO  - microPublication Biology
ER  -