---
_id: '9756'
abstract:
- lang: eng
  text: High-resolution visualization and quantification of membrane proteins contribute
    to the understanding of their functions and the roles they play in physiological
    and pathological conditions. Sodium dodecyl sulfate-digested freeze-fracture replica
    labeling (SDS-FRL) is a powerful electron microscopy method to study quantitatively
    the two-dimensional distribution of transmembrane proteins and their tightly associated
    proteins. During treatment with SDS, intracellular organelles and proteins not
    anchored to the replica are dissolved, whereas integral membrane proteins captured
    and stabilized by carbon/platinum deposition remain on the replica. Their intra-
    and extracellular domains become exposed on the surface of the replica, facilitating
    the accessibility of antibodies and, therefore, providing 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
    optimization of the SDS treatment to increase the labeling efficiency for quantification
    of Cav2.1, the alpha subunit of P/Q-type voltage-dependent calcium channels utilizing
    deep learning algorithms.
acknowledgement: This work was supported by the European Union (European Research
  Council Advanced grant no. 694539 and Human Brain Project Ref. 720270 to R. S.)
  and the Austrian Academy of Sciences (DOC fellowship to D.K.).
alternative_title:
- Neuromethods
article_processing_charge: No
author:
- first_name: Walter
  full_name: Kaufmann, Walter
  id: 3F99E422-F248-11E8-B48F-1D18A9856A87
  last_name: Kaufmann
  orcid: 0000-0001-9735-5315
- first_name: David
  full_name: Kleindienst, David
  id: 42E121A4-F248-11E8-B48F-1D18A9856A87
  last_name: Kleindienst
- 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: 'Kaufmann W, Kleindienst D, Harada H, Shigemoto R. High-Resolution localization
    and quantitation of membrane proteins by SDS-digested freeze-fracture replica
    labeling (SDS-FRL). In: <i> Receptor and Ion Channel Detection in the Brain</i>.
    Vol 169. Neuromethods. New York: Humana; 2021:267-283. doi:<a href="https://doi.org/10.1007/978-1-0716-1522-5_19">10.1007/978-1-0716-1522-5_19</a>'
  apa: 'Kaufmann, W., Kleindienst, D., Harada, H., &#38; Shigemoto, R. (2021). High-Resolution
    localization and quantitation of membrane proteins by SDS-digested freeze-fracture
    replica labeling (SDS-FRL). In <i> Receptor and Ion Channel Detection in the Brain</i>
    (Vol. 169, pp. 267–283). New York: Humana. <a href="https://doi.org/10.1007/978-1-0716-1522-5_19">https://doi.org/10.1007/978-1-0716-1522-5_19</a>'
  chicago: 'Kaufmann, Walter, David Kleindienst, Harumi Harada, and Ryuichi Shigemoto.
    “High-Resolution Localization and Quantitation of Membrane Proteins by SDS-Digested
    Freeze-Fracture Replica Labeling (SDS-FRL).” In <i> Receptor and Ion Channel Detection
    in the Brain</i>, 169:267–83. Neuromethods. New York: Humana, 2021. <a href="https://doi.org/10.1007/978-1-0716-1522-5_19">https://doi.org/10.1007/978-1-0716-1522-5_19</a>.'
  ieee: 'W. Kaufmann, D. Kleindienst, H. Harada, and R. Shigemoto, “High-Resolution
    localization and quantitation of membrane proteins by SDS-digested freeze-fracture
    replica labeling (SDS-FRL),” in <i> Receptor and Ion Channel Detection in the
    Brain</i>, vol. 169, New York: Humana, 2021, pp. 267–283.'
  ista: 'Kaufmann W, Kleindienst D, Harada H, Shigemoto R. 2021.High-Resolution localization
    and quantitation of membrane proteins by SDS-digested freeze-fracture replica
    labeling (SDS-FRL). In:  Receptor and Ion Channel Detection in the Brain. Neuromethods,
    vol. 169, 267–283.'
  mla: Kaufmann, Walter, et al. “High-Resolution Localization and Quantitation of
    Membrane Proteins by SDS-Digested Freeze-Fracture Replica Labeling (SDS-FRL).”
    <i> Receptor and Ion Channel Detection in the Brain</i>, vol. 169, Humana, 2021,
    pp. 267–83, doi:<a href="https://doi.org/10.1007/978-1-0716-1522-5_19">10.1007/978-1-0716-1522-5_19</a>.
  short: W. Kaufmann, D. Kleindienst, H. Harada, R. Shigemoto, in:,  Receptor and
    Ion Channel Detection in the Brain, Humana, New York, 2021, pp. 267–283.
date_created: 2021-07-30T09:34:56Z
date_published: 2021-07-27T00:00:00Z
date_updated: 2024-03-25T23:30:16Z
day: '27'
ddc:
- '573'
department:
- _id: RySh
- _id: EM-Fac
doi: 10.1007/978-1-0716-1522-5_19
ec_funded: 1
has_accepted_license: '1'
intvolume: '       169'
keyword:
- 'Freeze-fracture replica: Deep learning'
- Immunogold labeling
- Integral membrane protein
- Electron microscopy
language:
- iso: eng
month: '07'
oa_version: None
page: 267-283
place: New York
project:
- _id: 25CA28EA-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '694539'
  name: 'In situ analysis of single channel subunit composition in neurons: physiological
    implication in synaptic plasticity and behaviour'
- _id: 25CBA828-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '720270'
  name: Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)
publication: ' Receptor and Ion Channel Detection in the Brain'
publication_identifier:
  eisbn:
  - '9781071615225'
  isbn:
  - '9781071615218'
publication_status: published
publisher: Humana
quality_controlled: '1'
related_material:
  record:
  - id: '9562'
    relation: dissertation_contains
    status: public
series_title: Neuromethods
status: public
title: High-Resolution localization and quantitation of membrane proteins by SDS-digested
  freeze-fracture replica labeling (SDS-FRL)
type: book_chapter
user_id: D865714E-FA4E-11E9-B85B-F5C5E5697425
volume: 169
year: '2021'
...