Technical Reference #1794

1794.

Lipase Maturation Factor LMF1; Membrane Topology and Interaction with Lipase Proteins in the Endoplasmic Reticulum Mark H. Doolittle; Saskia B. Neher; Osnat Ben-Zeev; Jo Ling-liao; Ciara M. Gallagher; Maryam Hosseini; Fen Yin; Howard Wong; Peter Walter; Miklos Peterfy, University of California Los Angeles Veterans Affairs Greater Los Angeles Healthcare System University of California San Francisco Cedars-Sinai Medical Center, Journal of Biological Chemistry, 284(1794), (2009)
Link To Paper

Materials & Methods:
The cld mutant cell line has been described previously (15); it represents a hepatocyte-derived cell line derived from 16–18-day fetal cld/cld mice immortalized by the SV40 T antigen. Both cld mutant and HEK293 cell lines were maintained at a split ratio of 1:20 in Dulbecco’s modified Eagle’s medium 10% fetal bovine serum. Transfection of the cld mutant cell line used Effectene transfection reagent (Qiagen) at a DNA to reagent ratio of 1:10 and HEK293 cells were transfected with FuGENE 6 transfection reagent (Roche Applied Science) according to manufacturer’s instructions. Transfection was initiated 24 h after plating and cells were harvested 24–48 h post-transfection.

Microscopic Technique
Confocal microscopy

Cell Type(s)
HeLa cells; cld mutant and HEK293 cell lines;