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A new reliable and highly specific monoclonal antibody to detect the C-terminal region of silencer of cytokine signaling 1.
- Source :
-
European journal of haematology [Eur J Haematol] 2021 Jul; Vol. 107 (1), pp. 74-80. Date of Electronic Publication: 2021 May 06. - Publication Year :
- 2021
-
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.<br />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 SOCS1 <subscript>WT</subscript> stably transfected HEK293T cells, lymphoma cell lines and lymphoid tissues.<br />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.<br />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.<br /> (© 2021 The Authors. European Journal of Haematology published by John Wiley & Sons Ltd.)
- Subjects :
- 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 metabolism
Transfection
Suppressor of Cytokine Signaling 1 Protein chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1600-0609
- Volume :
- 107
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- European journal of haematology
- Publication Type :
- Academic Journal
- Accession number :
- 33714214
- Full Text :
- https://doi.org/10.1111/ejh.13620