Recombinant Mouse B-lymphocyte antigen CD20 (Ms4a1) is produced through a yeast expression system, targeting amino acid residues 132-291. This partial protein comes with an N-terminal 6xHis-tag, which streamlines purification processes. SDS-PAGE analysis confirms purity levels exceeding 90%, suggesting this protein offers dependable quality for research applications. The product is designed exclusively for research purposes and should not be considered for therapeutic applications.
CD20 appears to be a cell surface molecule found mainly on B-lymphocytes. It seems to play an important role in B-cell development and differentiation. Research indicates CD20 may be involved in signal transduction pathways that control cell cycle progression and proliferation. Studies examining CD20 interactions and functions are likely crucial for advancing immunology and hematological research.
Potential Applications
Note: The applications listed below are based on what we know about this protein's biological functions, published research, and experience from experts in the field. However, we haven't fully tested all of these applications ourselves yet. We'd recommend running some preliminary tests first to make sure they work for your specific research goals.
Based on the provided information, recombinant mouse CD20 is produced in a yeast expression system as a partial fragment (132-291aa) with an N-terminal 6xHis-tag. CD20 is a transmembrane protein with four membrane-spanning domains, and its native function depends on proper folding, membrane integration, and potential post-translational modifications. The expression region (132-291aa) likely covers a portion of the extracellular loops and transmembrane domains but lacks the full N-terminal region (1-131aa), which may be critical for complete structural integrity. Yeast expression systems provide eukaryotic folding machinery that can support disulfide bond formation and glycosylation, but membrane proteins like CD20 often require lipid environments for correct folding and may not integrate properly in soluble expression. The N-terminal His-tag could potentially interfere with folding or function. No validation data (e.g., antibody binding assays, cellular activity tests) are provided. Therefore, while yeast expression increases the probability of correct folding compared to prokaryotic systems, the protein's folding status and bioactivity cannot be confirmed without experimental validation, and the partial nature raises concerns about functional integrity.
1. Anti-CD20 Antibody Development and Characterization
This application is suitable as antibody generation primarily relies on linear epitope recognition, which is independent of folding status. The high purity and defined fragment support consistent immunization and screening. However, if correctly folded, antibodies may recognize conformational epitopes relevant to native CD20; if misfolded, generated antibodies might not optimally bind the full-length, membrane-integrated CD20 in biological contexts, limiting utility for functional assays like flow cytometry.
2. Protein-Protein Interaction Studies
If the recombinant CD20 fragment is correctly folded and retains native structure, it could be used in pull-down assays to identify interaction partners, as the His-tag facilitates immobilization. However, if misfolded (likely due to the partial nature and lack of membrane context), interaction domains may be altered, leading to non-specific binding or failure to recognize genuine biological partners, compromising the validity of identified networks.
3. ELISA-Based Binding Assays
If properly folded, the His-tagged CD20 fragment can be used as a coating antigen in ELISA to study antibody binding or screen interactions, with the tag enabling oriented immobilization. However, if misfolded, the protein may not present native epitopes accurately, resulting in unreliable binding kinetics or false negatives in antibody screening assays.
4. Structural and Biochemical Characterization Studies
If correctly folded, the fragment is suitable for biophysical analyses like circular dichroism or dynamic light scattering to study domain properties. However, if misfolded, structural data would misrepresent the native CD20 architecture, and the partial nature limits insights into full-length protein behavior. The His-tag may also interfere with some structural measurements.
Final Recommendation & Action Plan
This yeast-expressed partial CD20 fragment requires validation before functional applications due to the challenges of expressing transmembrane proteins in soluble form and the lack of full-length context; the recommended action plan includes: first validating folding through biophysical methods (e.g., circular dichroism for secondary structure, size-exclusion chromatography for oligomeric state) and functional assays (e.g., binding tests with known anti-CD20 antibodies); if validation confirms proper folding, proceed with applications while noting the limitations of the partial fragment and potential tag interference; if misfolded, consider using full-length CD20 expressed in mammalian systems for membrane integration or obtain a commercially validated standard; for immediate use, antibody development can proceed but validate antibodies against native CD20 in B-cells, and avoid interaction or binding studies until folding is confirmed. Always include appropriate controls, such as full-length CD20 or cell-based assays, to ensure biological relevance.
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