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Mouse monoclonal antibodies are uniform antibodies derived from a single B-cell clone through hybridoma technology, representing one of the most widely used antibody formats in life science research and in vitro diagnostics. This service serves academic laboratories, biotechnology companies, diagnostic developers, and pharmaceutical research teams requiring high specificity, long-term stable production, and batch-to-batch consistency of monoclonal antibodies.
We provide comprehensive services from antigen preparation, immunization, fusion, screening, subcloning stabilization, to antibody purification and quality control.
| Antigen Type | Recommended Immunization Strategy |
|---|---|
| Soluble high-purity protein | Conventional adjuvant immunization, titer monitoring at 4–6 weeks |
| Hydrophobic/membrane protein | Recombinant extracellular domain, LNP reconstitution, or cell-overexpression immunization |
| Short peptide (<15 aa) | KLH/BSA conjugation immunization |
| Phospho-epitope | Parallel immunization with phospho- and non-phospho peptides + subtractive screening |
| Conserved/low-immunogenicity protein | Multiple mouse strains, CpG adjuvant, or knockout mouse immunization |
From Antigen Design to Clone Discovery
The early stages of monoclonal antibody development focus on generating a strong immune response and identifying promising antibody-producing clones.
Figure 1: Early-stage workflow for mouse monoclonal antibody discovery.
Once positive clones have been identified, the next step is screening and validation to determine which candidates meet the project's performance requirements.
Screening protocols are customized based on final application requirements, not fixed templates.
ELISA: Binding activity, cross-reactivity, pairing capability
Western blot: Specificity confirmation under denatured and reducing conditions
Flow cytometry: Native conformation recognition of cell surface antigens
IHC/IF: Localization capability in tissues or cells
Neutralization/blocking assays: Functional activity screening (receptor-ligand systems)
| Phase | Description | Duration |
|---|---|---|
| Primary screening | Supernatant testing for antigen binding at 10–14 days post-fusion | 3–5 days |
| Secondary screening | Functional or cross-reactivity validation of positive wells | 1–2 weeks |
| Stabilization screening | Confirmation of antibody secretion stability after subcloning through serial passages | 2–5 weeks |
| Parameter | Method | Acceptance Criteria (Example) |
|---|---|---|
| Purity | SDS-PAGE (reducing/non-reducing) | >90% |
| Concentration | A280 or BCA | Reported actual value |
| Endotoxin | LAL method | <1 EU/mg (can be reduced to <0.1) |
| Isotype | Antibody isotyping kit | IgG1/2a/2b/3, κ/λ |
| Functional validation | ELISA/flow cytometry/WB, etc. | Consistent with project goals |
Additional characterization such as affinity (SPR/BLI), thermal stability (DSF), aggregation (SEC) can be added upon request.
Production and Quality Control
Following clone selection, antibodies undergo production, purification, and quality testing to ensure consistency and performance.
Figure 2: Antibody production, quality control, and final project deliverables.
Comprehensive quality assessment helps ensure that the final antibody meets the specifications required for downstream research and assay development.
| Feature | Technical Implication |
|---|---|
| High specificity | Recognizes single epitope, reduces cross-reactivity risk |
| Clone stability | Hybridoma cell lines can be long-term cryopreserved and revived with consistent performance |
| Batch reproducibility | Suitable for diagnostic reagents and long-term longitudinal studies |
| Broad compatibility | Validated with ELISA, WB, FC, IHC, IF, neutralization, blocking platforms |
| Mature hybridoma workflow | Well-defined technical parameters and quality checkpoints for immunization, fusion, and screening |
Q: How to choose between mouse monoclonal and rabbit monoclonal antibodies?
Mouse monoclonal antibodies offer more stable hybridoma and are more thoroughly validated in diagnostic applications. Rabbit monoclonal antibodies typically provide higher affinity for low-abundance antigens or certain phospho-epitopes. The two are not substitutes for each other.
Q: Can you guarantee that the final antibody will work in flow cytometry or IHC?
We use conditions aligned with final applications (e.g., cell-expressed antigens or tissue sections) whenever possible during screening. However, cross-target guarantees cannot be made. Screening protocols and acceptance criteria can be established at the initial project phase.
Q: Can hybridomas lose antibody production during long-term passage?
Stable subcloning combined with regular cryopreservation of master cell banks significantly reduces the risk of drift. We provide master cell bank cryopreservation services.
Q: Can I use only part of the service (e.g., immunization and fusion only)?
Yes. Service scope can be segmented according to project needs, with module-based pricing.
We have accumulated immunization and screening expertise on membrane proteins, phospho-specific antibodies, and low-immunogenicity targets using the hybridoma platform. In addition to mouse monoclonal antibodies, we also provide rabbit monoclonal antibodies, functional antibody screening, and diagnostic antibody pairing services.
Whether you are developing a new antibody for a specific assay or need a reliable hybridoma workflow, we can help design a plan aligned with your research goals.
Contact us to discuss your monoclonal antibody development needs.
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