Skip to main content
SpringerLink
Log in

Recycling cultured cells for immunofluorescent labeling

  • Original Paper
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract.

A method to use sequential rounds of immunofluorescent labeling in cell cultures is presented. The method is based on the utilization of a non-liquid reducing agent, sodium dithionite, in conjunction with ionic or non-ionic detergents (SDS or TX100, respectively) at room temperature. This method preserves cell morphology and substrate antigenicity, and operates through the complete extraction of most primary and secondary antibodies. Using this protocol, the sequential immunolocalization of different proteins is possible, without signal interference with previous immunolabeling rounds. In addition, the method is also useful to recycle blotted membranes in immunoblots.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Author information

Authors and Affiliations

  1. Departamento de Bioquímica, Instituto de Investigaciones Biomédicas, CSIC, Facultad de Medicina, Universidad Autónoma de Madrid, Arturo Duperier 4, 28029 Madrid, Spain, , , , ,

    Jesús Espada

  2. Departamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, Canto Blanco, 28049 Madrid, Spain, , , , ,

    Angeles Juarranz, Angeles Villanueva, Magdalena Cañete, Iván Andrés & Juan C. Stockert

Authors
  1. Jesús Espada
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Angeles Juarranz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Angeles Villanueva
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Magdalena Cañete
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Iván Andrés
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Juan C. Stockert
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Electronic Publication

Rights and permissions

Reprints and permissions

About this article

Cite this article

Espada, J., Juarranz, A., Villanueva, A. et al. Recycling cultured cells for immunofluorescent labeling. Histochem Cell Biol 116, 41–47 (2001). https://doi.org/10.1007/s004180100290

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s004180100290

Search

Navigation