Skip to content
DraconiticResearch
Search Sources API
201,683 studies31 sourcesUpdated 40 min ago
Peer-reviewedCC BY 4.0STAR Protocols

Protocol for FLOWMIST to profile autoreactivity of mouse monoclonal and serum antibodies by flow cytometry and microscopy

  • Michael Ivanov1
  • Uzair Ansari1
  • Jan Wisniewski1
  • Christian T. Mayer1
  1. 1Experimental Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 10 Center Dr, Bethesda, MD 20892, USA
Read full textDOICite

Abstract

Autoantibodies recognizing surface-bound or intracellular self-antigens are a hallmark of many autoimmune diseases. Here, we present a protocol for FLOWMIST to profile the autoreactivity of mouse monoclonal and serum antibodies using primary mouse splenocytes. We describe steps for antibody staining, quantitative flow cytometric screening, and confocal microscopy to classify antibody-binding patterns. Although demonstrated using mouse splenocytes, the workflow can be readily adapted to different target cells, antibody samples, species, and secondary detection reagents.

For complete details on the use and execution of this protocol, please refer to Simpson et al.1 and Newen et al.2

Related

  1. Asymmetrical Glyco-Engineering of Monoclonal Antibodies Provides Insights into α-Gal Immunogenicity
    Gerlof P. Bosman, Anna Ehlers, Thomas Buckley +6
    bioRxivPreprintLicence not statedFull textMolecular biology & gene regulationPMC
  2. High-throughput affinity profiling of antibodies against soluble and native membrane targets by digital flow cytometry
    Y. Cheng, S. Barcy, B. Fujimoto +3
    bioRxivPreprintCC BY-NC-NDFull textBiophysicsbioRxivPMC
  3. Establishment of Immunochromatographic Test Strips for Porcine Epidemic Diarrhea Virus (PEDV) Antibody Detection Based on Monoclonal Antibodies Against S2 Protein
    Meng Zhong, Rongchao Liu, Ruiying Wang +7
    BiomoleculesPeer-reviewedCC BY 4.0General biochemistryCrossref
  4. De novo design of monoclonal and bispecific antibodies with OFAntibody
    Valhalla Team
    arXivPreprintCC BY-NC-SA 4.0Full textBiophysicsarXiv
  5. From background to foreground: secondary antibodies coupled to lipophilic ATTO dyes enable high-density membrane labeling in super-resolution and expansion microscopy
    Jim Dompierre, Silvia del Pozo Perera, Louis Hurson +3
    HALPreprintCC BY-NCFull textBiophysicsHAL

Cite

BibTeXRIS
BibTeX
@article{ivanov2026protocol,
  title = {{Protocol for FLOWMIST to profile autoreactivity of mouse monoclonal and serum antibodies by flow cytometry and microscopy}},
  author = {Michael Ivanov and Uzair Ansari and Jan Wisniewski and Christian T. Mayer},
  journal = {STAR Protocols},
  year = {2026},
  month = oct,
  doi = {10.1016/j.xpro.2026.104918},
  url = {https://doi.org/10.1016/j.xpro.2026.104918},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - Protocol for FLOWMIST to profile autoreactivity of mouse monoclonal and serum antibodies by flow cytometry and microscopy
AU  - Michael Ivanov
AU  - Uzair Ansari
AU  - Jan Wisniewski
AU  - Christian T. Mayer
PY  - 2026
DA  - 2026/10/28
JO  - STAR Protocols
DO  - 10.1016/j.xpro.2026.104918
UR  - https://doi.org/10.1016/j.xpro.2026.104918
AB  - Autoantibodies recognizing surface-bound or intracellular self-antigens are a hallmark of many autoimmune diseases. Here, we present a protocol for FLOWMIST to profile the autoreactivity of mouse monoclonal and serum antibodies using primary mouse splenocytes. We describe steps for antibody staining, quantitative flow cytometric screening, and confocal microscopy to classify antibody-binding patterns. Although demonstrated using mouse splenocytes, the workflow can be readily adapted to different target cells, antibody samples, species, and secondary detection reagents.  For complete details on the use and execution of this protocol, please refer to Simpson et al.1 and Newen et al.2
SN  - 2666-1667
ER  -