PMID: 9632744Jun 20, 1998Paper

Immunoelectronmicroscopy of soluble and membrane proteins with a sensitive postembedding method

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
J E MoreiraT S Reese

Abstract

The application of immunoelectronmicroscopy to soluble proteins is limited because soluble proteins can redistribute during fixation. Fixation may also adversely affect the recognition of proteins associated with membranes. We show here how displacements of soluble proteins can be prevented and antigen sensitivity improved by freeze-substitution immunocytochemistry. The usefulness of this method for soluble cytoplasmic proteins is demonstrated for the twitchin protein in Aplysia muscle and the kinesin motor proteins in squid giant axons, in which the sizes of various cytoplasmic pools of kinesins are estimated. The utility for membrane proteins present in small numbers of copies is demonstrated by labeling a glutamate receptor subunit in mouse cerebellar cortex and the ZO-1 protein in tight junctions between MDCK cells. Thus, freeze-substitution immunocytochemistry can show the native distribution of both soluble and membrane proteins labeled with polyclonal antibodies and, at the same time, can reveal structural features comparable to those in chemically fixed or osmium freeze-substituted samples.

References

Jul 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·M TaubG Sato
Jul 1, 1991·The Journal of Cell Biology·N HirokawaS T Brady
Jan 1, 1989·The Journal of Cell Biology·P C Bridgman, M E Dailey
Oct 1, 1989·The American Journal of Physiology·B R StevensonM S Mooseker
Aug 30, 1994·Proceedings of the National Academy of Sciences of the United States of America·W C ProbstK R Weiss
Dec 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·E L BearerT S Reese
Jul 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·S B AndrewsT S Reese

❮ Previous
Next ❯

Citations

Nov 14, 2000·Journal of Neuroscience Methods·T DellerM Frotscher
May 7, 1999·European Journal of Biochemistry·M P Sheetz
Mar 22, 2002·Molecular Biology of the Cell·Joseph A DeGiorgisElaine L Bearer
Nov 10, 2015·Ultrastructural Pathology·Suélen MerloJorge E Moreira
May 19, 2004·Insect Biochemistry and Molecular Biology·N MonesiM L Paçó Larson
Sep 25, 2014·Frontiers in Neuroanatomy·Michael FrotscherShanting Zhao
Jul 1, 2005·Physiological Reviews·Scott L Hooper, Jeffrey B Thuma
May 22, 2004·BioTechniques·Dennis Richard LaJeunesseKathryn Fromson
Aug 2, 2003·Biochemical and Biophysical Research Communications·Luciana CasalettiRoy E Larson

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.