---
OA_type: closed access
_id: '19990'
abstract:
- lang: eng
  text: Visualizing molecular localization at high resolution contributes to understanding
    of their functions and roles in physiological and pathological conditions. Sodium
    dodecyl sulfate-digested freeze-fracture replica labeling (SDS-FRL) is a powerful
    electron microscopy method to study high-resolution two-dimensional distribution
    of transmembrane proteins and their tightly associated proteins on platinum-carbon
    replica. During treatment with SDS, unfixed proteins and intracellular organelle
    are dissolved and integral membrane proteins captured and stabilized by carbon
    and platinum deposition are denatured, retaining most of their antigenicity, and
    exposed on exoplasmic and protoplasmic surfaces of lipid monolayers. The exposure
    of these antigens on the surface of replica facilitates the accessibility of antibodies
    and therefore provides higher labeling efficiency than those obtained with other
    immunoelectron microscopy techniques. In this chapter, we describe the protocols
    of SDS-FRL adapted for mammalian brain samples and an additional procedure for
    fluorescence-guided electron microscopy for replica immunolabeling.
acknowledgement: We thank Mitsuru Ikeda for preparing replica images used in Fig.
  2.
article_processing_charge: No
author:
- first_name: Harumi
  full_name: Harada, Harumi
  id: 2E55CDF2-F248-11E8-B48F-1D18A9856A87
  last_name: Harada
  orcid: 0000-0001-7429-7896
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
citation:
  ama: 'Harada H, Shigemoto R. High-Resolution Localization of Membrane Proteins by
    SDS-Digested Freeze-Fracture Replica Labeling (SDS-FRL). In: <i>Receptor and Ion
    Channel Detection in the Brain</i>. Neuromethods. Springer Nature; 2016:233-245.
    doi:<a href="https://doi.org/10.1007/978-1-4939-3064-7_17">10.1007/978-1-4939-3064-7_17</a>'
  apa: Harada, H., &#38; Shigemoto, R. (2016). High-Resolution Localization of Membrane
    Proteins by SDS-Digested Freeze-Fracture Replica Labeling (SDS-FRL). In <i>Receptor
    and Ion Channel Detection in the Brain</i> (pp. 233–245). Springer Nature. <a
    href="https://doi.org/10.1007/978-1-4939-3064-7_17">https://doi.org/10.1007/978-1-4939-3064-7_17</a>
  chicago: Harada, Harumi, and Ryuichi Shigemoto. “High-Resolution Localization of
    Membrane Proteins by SDS-Digested Freeze-Fracture Replica Labeling (SDS-FRL).”
    In <i>Receptor and Ion Channel Detection in the Brain</i>, 233–45. Neuromethods.
    Springer Nature, 2016. <a href="https://doi.org/10.1007/978-1-4939-3064-7_17">https://doi.org/10.1007/978-1-4939-3064-7_17</a>.
  ieee: H. Harada and R. Shigemoto, “High-Resolution Localization of Membrane Proteins
    by SDS-Digested Freeze-Fracture Replica Labeling (SDS-FRL),” in <i>Receptor and
    Ion Channel Detection in the Brain</i>, Springer Nature, 2016, pp. 233–245.
  ista: 'Harada H, Shigemoto R. 2016.High-Resolution Localization of Membrane Proteins
    by SDS-Digested Freeze-Fracture Replica Labeling (SDS-FRL). In: Receptor and Ion
    Channel Detection in the Brain. , 233–245.'
  mla: Harada, Harumi, and Ryuichi Shigemoto. “High-Resolution Localization of Membrane
    Proteins by SDS-Digested Freeze-Fracture Replica Labeling (SDS-FRL).” <i>Receptor
    and Ion Channel Detection in the Brain</i>, Springer Nature, 2016, pp. 233–45,
    doi:<a href="https://doi.org/10.1007/978-1-4939-3064-7_17">10.1007/978-1-4939-3064-7_17</a>.
  short: H. Harada, R. Shigemoto, in:, Receptor and Ion Channel Detection in the Brain,
    Springer Nature, 2016, pp. 233–245.
corr_author: '1'
date_created: 2025-07-10T13:56:06Z
date_published: 2016-02-02T00:00:00Z
date_updated: 2026-04-07T08:32:03Z
day: '02'
department:
- _id: RySh
doi: 10.1007/978-1-4939-3064-7_17
language:
- iso: eng
month: '02'
oa_version: None
page: 233-245
publication: Receptor and Ion Channel Detection in the Brain
publication_identifier:
  eisbn:
  - '9781493930647'
  eissn:
  - 1940-6045
  isbn:
  - '9781493930630'
  issn:
  - 0893-2336
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
series_title: Neuromethods
status: public
title: High-Resolution Localization of Membrane Proteins by SDS-Digested Freeze-Fracture
  Replica Labeling (SDS-FRL)
type: book_chapter
user_id: ba8df636-2132-11f1-aed0-ed93e2281fdd
year: '2016'
...