Choosing the right antibody for resistin-like molecule (RELM/FIZZ) family members

Histochem Cell Biol. 2013 Apr;139(4):605-13. doi: 10.1007/s00418-012-1042-0. Epub 2012 Oct 18.

Abstract

The family of resistin-like molecules (RELM), also known as found in inflammatory zone (FIZZ), consists of four members in mouse (RELMα/FIZZ1/HIMF, RELMβ/FIZZ2, Resistin/FIZZ3, and RELMγ/FIZZ4) and two members in human (resistin and RELMβ). The importance of these proteins in many aspects of physiology and pathophysiology, especially inflammatory processes, is rapidly evolving in the literature, and many investigators are beginning to work in this field. Most published studies focus on only one isoform, do not evaluate other isoforms that might be present, and have not tested for the specificity of the antibody used. Because RELM isoforms have high sequence and structural similarity and both distinct and overlapping functions, it is important to use a specific antibody to distinguish each isoform in the study. We constructed and established HEK 293 cell lines that constitutively express each isoform. Using these cell lines, we determined the specificity of antibodies (both commercially available and laboratory-made) to each isoform by Western blot and immunofluorescence. Some of the antibodies showed specificity in Western blotting but were not applicable in immunofluorescence. Others showed cross reactivity in Western blot assays. Our results indicate that RELM antibody specificity should be taken into account when using them in research and interpreting data obtained with them.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibodies / immunology*
  • Antibody Specificity
  • Cell Line
  • HEK293 Cells
  • Hormones, Ectopic / immunology*
  • Humans
  • Intercellular Signaling Peptides and Proteins / immunology*
  • Mice
  • Protein Isoforms / immunology
  • Recombinant Fusion Proteins / immunology
  • Resistin / immunology*

Substances

  • Antibodies
  • Hormones, Ectopic
  • Intercellular Signaling Peptides and Proteins
  • Protein Isoforms
  • RETNLB protein, human
  • Recombinant Fusion Proteins
  • Resistin
  • Retnla protein, mouse
  • Retnlb protein, mouse
  • resistin-like gamma, mouse