TY - JOUR
T1 - Transmission Electron Microscopy and Immunocytochemical Studies of Yeast
T2 - Analysis of HMG—CoA Reductase Overproduction by Electron Microscopy
AU - Wright, Robin
AU - Rine, Jasper
PY - 1989/1/1
Y1 - 1989/1/1
N2 - Immunocytochemistry provides a powerful technique for analysis of molecular organization within cells. Using specific antibody probes together with electron microscopy (EM), the localization of a protein within individual cells can be analyzed. During the electronmicroscopic analysis of yeast cells, it is found that samples prepared for routine EM could be labeled quite successfully with affinity-purified antiserum against the membrane protein, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA reductase), and colloidal gold reagents. This chapter describes the detailed protocols of these methods. Generation of reagent antibodies of high specificity is of utmost importance for accurate immunolocalization at the EM level. The electron micrographs presented in the chapter compare several fixation protocols and resins, and the effect of these variables on immunoreactivity of HMG-CoA reductase with affinity-purified antiserum.
AB - Immunocytochemistry provides a powerful technique for analysis of molecular organization within cells. Using specific antibody probes together with electron microscopy (EM), the localization of a protein within individual cells can be analyzed. During the electronmicroscopic analysis of yeast cells, it is found that samples prepared for routine EM could be labeled quite successfully with affinity-purified antiserum against the membrane protein, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA reductase), and colloidal gold reagents. This chapter describes the detailed protocols of these methods. Generation of reagent antibodies of high specificity is of utmost importance for accurate immunolocalization at the EM level. The electron micrographs presented in the chapter compare several fixation protocols and resins, and the effect of these variables on immunoreactivity of HMG-CoA reductase with affinity-purified antiserum.
UR - http://www.scopus.com/inward/record.url?scp=0024388570&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0024388570&partnerID=8YFLogxK
U2 - 10.1016/S0091-679X(08)61624-6
DO - 10.1016/S0091-679X(08)61624-6
M3 - Article
C2 - 2674630
AN - SCOPUS:0024388570
VL - 31
SP - 473
EP - 512
JO - Methods in Cell Biology
JF - Methods in Cell Biology
SN - 0091-679X
IS - C
ER -