Lab protocols
Heather Harding's protocol for detection of PERK or GCN2 protein in cell lysates or tissue extracts by combined immunoprecipitation western immunobloting (download PDF)
Heather Harding's protocol for detection of phosphorylated eIF2α by western blotting (download pdf)
Heather Harding's protocol for isolating nuclear and cytoplasmic extracts
Culturing cells with defective ISR
Anne Bertolotti's protocol for culture of AR42J cells
Heather Harding's protocol for immortalizing MEFs with SV-40 large T antigen
Heather Hardings protocol for producing VSV-G pseudotyped retroviruses
How to obtain full-length cDNA clones
Steffen Preissler's protocol for native gels
Protocols related to mutant mice
Whilst we no longer maintain stocks of live mice, most of the strains made by the lab over the years have been cryopreserved as embryos and are available for sharing. Shipping frozen mouse embryos is an expensive undertaking and must be given due consideration. Below is a list of genotyping protocols for alleles created in the lab. They are available upon request
Detection of AIRAP genotype in knockout mice
Detection of Alb::A1 (A.K.A. Alb::GADD34(+) transgene)
Detection of ATF4 genotype (Leiden's mutation) and wildtype>
Detection of CHOP genotype in knockout mice
Detection of CRE in transgenic mice
ERO1β mutant mice
Detection of GADD34 genotype in knockout mice (WT & KO1 Ex allele)
Detection of GCN2 genotype in KO4 knockout mice and cells
Detection of IRE1aα genotype in knockout mice and cell lines
Detection of IRE1β genotype in knockout mice
Detection of PERK genotype in knockout mice and cell lines