Anti-Human CD20 [Clone 2H7] — Purified in vivo GOLD™ Functional Grade

The clone 2H7, a mouse anti-human CD20 antibody, is primarily used to recognize and bind to the CD20 antigen on human B cells. While specific details on its use in in vivo mouse studies might not be directly stated, researchers could potentially use this antibody in models where human B cells are present, such as humanized mice. Humanized mice are engineered to carry human immune cells, including B cells, which could be targeted by the 2H7 antibody for studying B-cell biology or diseases like B-cell lymphomas.

In studies involving humanized mice, the 2H7 antibody could be employed to:

1. Deplete or track human B cells: By binding to CD20 on human B cells, the 2H7 antibody could be used to study B-cell behavior or to deplete B cells in specific experimental conditions.

2. Study B-cell mediated diseases: Since CD20 is a target for treatments of B-cell diseases, the 2H7 antibody might be used to simulate or study therapeutic interventions in humanized mouse models.

3. Examine B-cell development and function: The presence of CD20 on pre-B cells and mature B cells makes the 2H7 antibody useful for investigating B-cell development stages in humanized mice.

However, for direct in vivo studies in mice, researchers would need to ensure that the antibody does not cross-react with mouse CD20 (which is not specified in the provided sources) or that the mouse model contains human B cells that express CD20.

The correct storage temperature for sterile packaged clone 2H7 (anti-human CD20 antibody) is 2°C to 8°C (approximately 36°F to 46°F).

Storage Guidelines for Clone 2H7

Recommended Temperature Range: All sources consistently specify that this antibody should be stored between 2°C and 8°C. This refrigerated temperature range maintains the antibody's stability and binding capacity while preventing degradation.

Additional Storage Considerations: The antibody should be protected from prolonged exposure to light and must not be frozen. For optimal storage conditions, keep the antibody solution undiluted in its original formulation.

Alternative Storage Options

For different storage durations, Bio-Rad recommends aliquoting the antibody and storing at -20°C upon receipt. However, this appears to be specific to their preparation and packaging, while the standard sterile packaged versions consistently specify the 2-8°C range.

The phosphate-buffered solution formulation with 0.09% sodium azide preservative helps maintain stability at the recommended refrigerated temperature. This preservative system is specifically designed to prevent microbial growth while maintaining antibody integrity during storage.

Some of the most commonly used antibodies or proteins alongside 2H7 in the literature include rituximab, F8, and isotype control antibodies. The choice depends on the antigen, experimental design, and biological application. Below are context-specific details according to distinct uses of 2H7:

• Anti-CD20 antibodies:
  2H7 has been frequently compared or used with rituximab, another monoclonal antibody targeting the CD20 antigen on B cells. Both antibodies bind to overlapping epitopes on CD20, but structural analyses reveal differences in affinity and binding interactions. Rituximab is the clinically dominant anti-CD20 antibody and serves as a standard control or reference in studies evaluating 2H7's binding and functional characteristics.

• Isotype controls:
  In functional assays, particularly opsonophagocytic and cytotoxicity tests, 2H7 is commonly paired with an isotype control antibody of the same subclass (e.g., IgG1, IgG2b) but irrelevant specificity, to confirm specificity of effect. For example, in the context of S. aureus infection models, 2H7 was directly compared to an IgG1 control antibody in bacterial killing assays.

• F8 antibody (Anti-EDA domain of fibronectin):
  In tumor targeting and antibody engineering research, 2H7 (targeting a unique epitope on the EDA domain) has been used in combination with the F8 antibody, which recognizes a distinct but neighboring epitope on the same protein. Bispecific constructs (chelated recombinant antibodies, CRAbs) combining F8 and 2H7 have been evaluated for their biodistribution and specificity in tumor models.

• Antibodies targeting different domains of the same antigen:
  In studies involving ELISA and epitope mapping, 2H7 is used alongside antibodies recognizing alternative domains or recombinant fragments of target proteins, such as SasA fragments (SRR1, NRR1, NRR, NRR2). This allows detailed characterization of epitope specificity.

• Other monoclonal antibodies recognizing different antigens for multiplex assays:
  2H7, especially when used for phenotyping or protein targeting, may be included with panels of other mAbs (such as anti-CD19, anti-CD22 for B cells, or anti-CD3 for T cells), though specific combinations depend on the experimental design and are less frequently reported in direct conjunction with 2H7 in published literature.

Summary table:

These are the most commonly documented antibodies/proteins paired with 2H7 for comparative, control, or multi-targeting purposes in published biomedical research.

Clone 2H7 is a mouse monoclonal antibody that specifically recognizes the human CD20 cell surface antigen—a 33-37 kDa non-glycosylated phosphoprotein present on normal and malignant B cells. Key findings from scientific literature using clone 2H7 can be summarized as follows:

• Specificity and Epitope Recognition: Clone 2H7 binds to a defined sequence (YNCEPANPSEKNSPST) in the large extracellular loop of human CD20. It only recognizes human CD20 in its native oligomeric form, meaning the antibody binds to CD20 when it is naturally assembled on the cell surface, not to denatured or fragmented protein.

• Expression Profile of CD20: CD20 is expressed throughout pre-B cell development and on mature B cells—resting or activated. It is lost upon differentiation into plasma cells, which clarifies its utility for targeting B cells in diagnostics and therapeutics without affecting plasma cells.

• Therapeutic Implications and Engineering: The original 2H7 is an IgG2b isotype, unable to mediate effective antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC) with human cells or serum. A chimeric version of 2H7 was developed by replacing its constant domains with human gamma 1 and kappa domains, retaining the same binding specificity but gaining strong ADCC and CDC activity—making it significantly more potent for therapeutic applications, especially for B-cell lymphomas.

• Applications: Clone 2H7 is widely used in flow cytometry and immunophenotyping to identify and quantify B cells in research, clinical immunology, and oncology settings. The antibody is also commonly applied in the development and validation of anti-CD20 therapies.

• Cross Reactivity: Clone 2H7 primarily recognizes human CD20 but may have some reactivity with rhesus monkey CD20, indicating potential use in specific non-human primate studies.

These findings affirm clone 2H7’s central role in B-cell biology research, therapeutic antibody engineering, and clinical diagnostics related to CD20.