Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Mapping the dialogue: Decoding alveolar stem-niche interactions.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

High-throughput histopathology for complex in vitro models.

Cell reports methods·2026
Same author

Ubiquitin Ligase COP1 Suppresses Neuroinflammation by Degrading c/EBPβ in Microglia.

Cell·2026
Same author

cFLIP suppresses caspase-1- and MLKL-independent perinatal lethality driven by auto-processing impaired caspase-8 D387A.

Cell death and differentiation·2025
Same author

Author Correction: Activity of caspase-8 determines plasticity between cell death pathways.

Nature·2025
Same author

Author Correction: Cleavage of RIPK1 by caspase-8 is crucial for limiting apoptosis and necroptosis.

Nature·2025

Related Experiment Video

Updated: Sep 1, 2025

Standardized Processing for Formalin-Fixed, Paraffin-Embedded Cell Pellet Immunohistochemistry Controls
06:43

Standardized Processing for Formalin-Fixed, Paraffin-Embedded Cell Pellet Immunohistochemistry Controls

Published on: July 27, 2022

8.1K

Standardized Processing for Formalin-Fixed, Paraffin-Embedded Cell Pellet Immunohistochemistry Controls.

Charles Havnar1, Kathy Hotzel1, Carmina Espiritu1

  • 1Department of Pathology, Genentech.

Journal of Visualized Experiments : Jove
|August 15, 2022
PubMed
Summary

Developing reliable immunohistochemistry (IHC) assays requires effective controls. This study presents a protocol for creating formalin-fixed, paraffin-embedded cell pellet controls for IHC method development, especially for novel proteins.

More Related Videos

Determination of Protein Expression Level in Cultured Cells by Immunocytochemistry on Paraffin-embedded Cell Blocks
09:06

Determination of Protein Expression Level in Cultured Cells by Immunocytochemistry on Paraffin-embedded Cell Blocks

Published on: May 20, 2018

11.6K
Synthetic Antigen Controls for Immunohistochemistry
09:30

Synthetic Antigen Controls for Immunohistochemistry

Published on: August 23, 2021

2.6K

Related Experiment Videos

Last Updated: Sep 1, 2025

Standardized Processing for Formalin-Fixed, Paraffin-Embedded Cell Pellet Immunohistochemistry Controls
06:43

Standardized Processing for Formalin-Fixed, Paraffin-Embedded Cell Pellet Immunohistochemistry Controls

Published on: July 27, 2022

8.1K
Determination of Protein Expression Level in Cultured Cells by Immunocytochemistry on Paraffin-embedded Cell Blocks
09:06

Determination of Protein Expression Level in Cultured Cells by Immunocytochemistry on Paraffin-embedded Cell Blocks

Published on: May 20, 2018

11.6K
Synthetic Antigen Controls for Immunohistochemistry
09:30

Synthetic Antigen Controls for Immunohistochemistry

Published on: August 23, 2021

2.6K

Area of Science:

  • Biomedical Sciences
  • Cell Biology
  • Histotechnology

Background:

  • Positive and negative controls are crucial for immunohistochemistry (IHC) assay development.
  • Tissue controls are limited for novel or ubiquitously expressed proteins.
  • Cell pellets offer standardized controls for antibody characterization.

Purpose of the Study:

  • To describe a protocol for creating formalin-fixed, paraffin-embedded (FFPE) cell pellet controls.
  • To provide a method for IHC assay development, particularly for novel proteins.
  • To establish standardized controls for antibody selection and validation.

Main Methods:

  • Processing and embedding cell pellets to mimic tissue processing.
  • Utilizing cell lines with defined protein/transcript expression levels (high, medium, low).
  • Incorporating engineered cell lines (over-expressing or gene-deleted via CRISPR).

Main Results:

  • Successfully created FFPE cell pellet controls.
  • Demonstrated the utility of cell pellets for IHC method development.
  • Validated the recapitulation of tissue processing procedures.

Conclusions:

  • FFPE cell pellet controls are valuable for IHC assay development, especially for challenging targets.
  • This protocol facilitates the creation of standardized controls for antibody validation.
  • Cellular controls enhance the reliability of IHC assays for research and diagnostics.