Measurement of labile and protein-bound heme in fixed prostate cancer cells and in cellular fractions

STAR Protoc. 2021 Apr 22;2(2):100491. doi: 10.1016/j.xpro.2021.100491. eCollection 2021 Jun 18.

Abstract

Labile heme is present in the cells at very low concentrations, either unbound or loosely bound to molecules, and accessible for signaling as alarmin. Our recent work suggests that extracellular heme can be taken up and detected in the nuclei of cancer cells. Here, we describe the detailed protocol for detection of labile and total heme in prostate cancer cells and its measurement in subcellular compartments in vitro. The protocol can be adapted to be used for other cell types. For complete details on the use and execution of this protocol, please refer to Canesin et al. (2020).

Keywords: Cancer; Cell Biology; Cell separation/fractionation; Cell-based Assays; Protein Biochemistry.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques
  • Cell Fractionation / methods*
  • Cell Line, Tumor
  • Cells, Immobilized / chemistry
  • Colorimetry / methods*
  • Heme / analysis*
  • Humans
  • Male
  • Prostatic Neoplasms* / chemistry
  • Prostatic Neoplasms* / metabolism

Substances

  • Heme