ERp72: Lysates

ERp72 is an endoplasmic reticulum luminal protein that is both a stress protein and a member of the protein disulfide isomerase family of proteins. Sequence analysis of ERp72 shows that it shares sequence identity with PDI (protein disulphide isomerase) at three discrete regions, as well as containing the ER retention signal KEEL at its C-terminus. ERp72 is shown to have three copies of sequence believed to be CGHC-containing active sites, which are also found in PDI. These CGHC sites are critical sites that are involved in the protein disulphide isomerization process. These results suggest that ERp72 is a newly described member of the family of CGHC-containing proteins that functions as a molecular chaperone. This antibody was raised by a genetic immunization technique. Genetic immunization can be used to generate antibodies by directly delivering antigen-coding DNA into the animal, rather than injecting a protein or peptide (Tang et al. PubMed: 1545867; Chambers and Johnston PubMed 12910245; Barry and Johnston PubMed: 9234514). The animal's cells produce the protein, which stimulates the animal's immune system to produce antibodies against that particular protein. A vector coding for a partial fusion protein was used for genetic immunisation of a mouse and the resulting serum was tested in Western blot against an E.coli lysate containing that partial fusion protein. Genetic immunization offers enormous advantages over the traditional protein-based immunization method. DNA is faster, cheaper and easier to produce and can be produced by standard techniques readily amenable to automation. Furthermore, the antibodies generated by genetic immunization are usually of superior quality with regard to specificity, affinity and recognizing the native protein.