A new reliable and highly specific monoclonal antibody to detect the C-terminal region of silencer of cytokine signaling 1.

Abstract

INTRODUCTION:

SOCS1, a negative regulator of JAK/STAT signaling, is among the most frequently mutated genes in DLBCL and classical Hodgkin lymphoma. The C-terminal SOCS box domain, mediating the degradation of phospho-JAK2, is often affected or even lacking. The analysis of such variants is hampered by the lack of a SOCS1-specific monoclonal antibody recognizing the C-terminus of SOCS1. As this C-terminus is often lost or mutated in B-cell lymphomas, staining with amino-terminal targeting antibodies in a lymphoma setting might be misleading.

METHODS:

BALB/c mice were immunized with a truncated SOCS1 C-terminal protein. The supernatant of generated hybridoma cells was screened by ELISA and, immunohistochemically, on formalin-fixed and paraffin-embedded tonsil. After antibody purification by affinity chromatography, epitope mapping and cross-reactivity check followed via substitution scans. SOCS1 protein expression was investigated on cell cultures and cytoblocks of SOCS1WT stably transfected HEK293T cells, lymphoma cell lines and lymphoid tissues.

RESULTS:

Procedures resulted in one monoclonal IgG1 anti-SOCS1 antibody, 424C, that recognizes and strongly binds to the C-terminal region of SOCS1 in immunoblot and immunohistochemistry analyses.

CONCLUSION:

This new anti-SOCS1 monoclonal antibody is a valuable tool to detect SOCS1 expression dependent on an existing SOCS1 box and, therefore, indicating a full-length SOCS1 protein.

• Authors: Stephanie E Weissinger, Malena Zahn, Ralf Marienfeld, Claudia Tessmer, Gerhard Moldenhauer, Peter Möller
• Journal: European journal of haematology (Eur J Haematol) Vol. 107 Issue 1 Pg. 74-80 (Jul 2021) ISSN: 1600-0609 [Electronic] England
• PMID: 33714214 (Publication Type: Journal Article)
• Copyright: © 2021 The Authors. European Journal of Haematology published by John Wiley & Sons Ltd.

Chemical References

• Antibodies, Monoclonal
• Epitopes
• SOCS1 protein, human
• Suppressor of Cytokine Signaling 1 Protein

Topics

• Animals
• Antibodies, Monoclonal (chemistry)
• Binding Sites
• Epitope Mapping
• Epitopes (chemistry)
• HEK293 Cells
• Humans
• Hybridomas (metabolism)
• Lymphoid Tissue (metabolism)
• Lymphoma (metabolism)
• Lymphoma, B-Cell (genetics)
• Mice
• Mice, Inbred BALB C
• Mutation
• Palatine Tonsil (metabolism)
• Protein Domains
• Signal Transduction
• Suppressor of Cytokine Signaling 1 Protein (chemistry, metabolism)
• Transfection