Anti-Human CD2 [Clone G11] — Purified in vivo GOLD™ Functional Grade

Clone G11 is most commonly referenced in vivo mouse studies as an EGFP-labeled mouse embryonic stem cell (mESC) line that facilitates cell tracking and transplantation studies. EGFP (Enhanced Green Fluorescent Protein) allows for direct visualization of the fate, integration, and differentiation of transplanted cells within recipient mice without additional labeling steps.

Key in vivo uses and context for mESC clone G11:

• Generation of Chimeric Mice: EGFP-mESC clone G11 can contribute to multiple tissues in chimeric mice, demonstrating pluripotency by differentiating into derivatives of all three germ layers (ectoderm, mesoderm, and endoderm) both in vitro and in vivo.
• Cell Fate and Lineage Tracing: High levels of EGFP expression are maintained after differentiation, enabling researchers to trace donor cells after engraftment or transplantation—even after prolonged periods (e.g., 12 weeks post-transplant in neural tissue).
• Transplantation and Tissue Replacement: EGFP-mESC(G11) cells have been differentiated into early neuronal precursors and subsequently transplanted into syngeneic mouse brains, with long-term survival and identification of the transplanted cells.

Advantages of G11 for in vivo studies:

• Avoids potential artifacts associated with chemical or viral labeling of ESCs.
• Cells can be robustly traced by EGFP fluorescence after transplantation and differentiation, supporting detailed analyses in cell and tissue replacement/regeneration experiments.

Alternative uses or confusion:There exist other "G11" clones in the literature (such as antibody clones), but in the context of in vivo mouse studies, the EGFP-mESC clone G11 is the recognized reference. If you were seeking information about the use of a G11 antibody (e.g., against a specific antigen in vivo), please clarify the target or species specificity.

Summary Table: Key Applications of EGFP-mESC Clone G11 in Mouse Studies

If a different G11 antibody clone was intended (e.g., against tenascin-C for tumor targeting), please specify for a tailored response.

Based on the available information, the Lambda gt11 vector should be stored at -20°C. This temperature recommendation comes directly from the storage conditions specified for the Lambda gt11 Vector Kit.

Storage Requirements for Lambda gt11

The Lambda gt11 expression vector, which is used for constructing cDNA libraries, has specific storage requirements to maintain its viability and functionality. The vector can accept inserts up to 7.2 kilobases in length and is designed for creating recombinant phage that can be identified through blue-white color screening.

Additional Storage Considerations

For optimal preservation of the Lambda gt11 system, other components have different storage requirements:

• Bacterial glycerol stocks associated with the system can be stored at either -20°C or -80°C
• Packaging extracts (like Gigapack III) require storage at -80°C and are extremely temperature-sensitive
• Control DNA should be stored at -80°C immediately upon receipt

Important Storage Guidelines

The storage instructions emphasize avoiding repeated freeze-thaw cycles, with a maximum of two freeze-thaw cycles recommended. For short-term storage needs, some components can be kept at 4°C for up to one month, but the primary vector should maintain the -20°C storage temperature for long-term stability.

The -20°C storage temperature for the Lambda gt11 vector ensures maximum viability and prevents degradation that could affect cloning efficiency and experimental results.

Antibodies and Proteins Commonly Used with G11 in the Literature

G11 most often refers to a human monoclonal antibody (scFv G11) that selectively targets the extra-domain C of tenascin-C, a protein highly expressed in solid tumors such as lung cancer and mesothelioma. In the literature, G11 is frequently used in combination with other proteins, antibody formats, or functional domains to enhance tumor targeting, therapeutic delivery, or imaging. Here are the most notable examples:

Common Protein and Antibody Partners for G11

• Interleukin-2 (IL-2): The G11 antibody has been genetically fused to human interleukin-2 (scFv(G11)-IL2) to create an immunocytokine. This fusion aims to deliver the cytokine directly to the tumor microenvironment, potentially enhancing local immune activation against cancer cells. This format is compared to similar fusions with the L19 antibody (scFv(L19)-IL2), which forms non-covalent homodimers, whereas the G11-IL2 fusion is monomeric under physiological conditions.
• Small Immunoprotein (SIP) Format: The G11 scFv has been cloned into a SIP format by fusing it to the human ?CH4 domain of the secretory isoform S2 of human IgE. This results in a 75 kDa bivalent miniantibody (SIP(G11)), which has been shown to offer advantages over IgG and standard scFv formats, particularly for radioimmunotherapy applications.
• Tumor Penetrating Peptide iRGD: The scFv G11 has also been functionalized with the tumor-penetrating peptide iRGD. This modification improves tumor homing, extravasation, and penetration into the tumor parenchyma, as demonstrated in U87-MG mouse models.

Related Therapeutic Contexts

While the above combinations are the most directly described in the literature for G11, the broader field of antibody-based therapeutics often involves:

• Cytotoxic Payloads: Antibodies like G11 are candidates for conjugation with radioactive isotopes or cytotoxic drugs (antibody-drug conjugates, ADCs), though such specific conjugates for G11 are not detailed in the provided literature.
• Cytokine Fusions: Beyond IL-2, other cytokines (e.g., IL-12, TNF) are often explored in the context of tumor-targeting antibodies, though only IL-2 fusion is specifically mentioned with G11.
• Bispecific Antibodies: While not explicitly described for G11, bispecific formats (targeting two different antigens) are a common strategy in oncology, and could theoretically be explored with G11 in future research.

Summary Table

Conclusion

The most commonly used proteins and antibody formats with G11 in the literature are interleukin-2 (for immunocytokine therapy), the SIP miniantibody format (for improved tumor targeting), and the iRGD tumor-penetrating peptide (for enhanced tumor parenchyma penetration). These examples illustrate how G11 serves as a versatile scaffold for constructing advanced tumor-targeting biologics.

Key Findings from Clone G11 Citations

The search results reference two distinct "clone G11" antibodies in immunological research, each with specific properties and findings. Here, we summarize the principal findings for each:

Clone G11-6 (Anti-canine/feline PD-L1 Monoclonal Antibody)

• Cross-Species Reactivity: Clone G11-6, originally identified as an anti-canine PD-L1 monoclonal antibody, was shown to bind feline PD-L1 (fPD-1) by flow cytometry using NIH3T3/fPDL1 cells.
• Epitope Specificity: This antibody seems to recognize a conformational epitope of feline PD-L1, as it could not detect the protein by western blotting unless the protein was first immunoprecipitated, suggesting the epitope is dependent on protein folding or structure rather than linear sequence.
• Diagnostic Utility: Flow cytometry with clone G11-6 detected PD-L1 expression in specific feline cancer cell lines—three mammary adenocarcinomas, a T-cell lymphoma, and a B-cell lymphoma—but not in macrophage or fibroblast cell lines. This suggests selective expression of PD-L1 in certain feline malignancies, potentially useful for diagnostic or research applications.
• Potential Therapeutic Implications: Although primarily used for detection, the identification of PD-L1-expressing feline tumors may have implications for immunotherapy research, including checkpoint inhibitor studies in veterinary oncology.

Clone scFv(G11) (Human Anti-Tenascin-C Monoclonal Antibody Fragment)

• High-Affinity Binding: In a study of human monoclonal antibodies targeting domain C of tenascin-C, the single-chain variable fragment (scFv) clone G11 was isolated for its slowest kinetic dissociation from the antigen. This implies that clone G11 has a high binding affinity for its target.
• Purification and Stability: The study reported that this clone was successfully purified, indicating it can be produced in quantity and is stable enough for further biochemical characterization.
• Potential Applications: High-affinity, slow-dissociating antibodies like scFv(G11) are valuable for diagnostic imaging, targeted therapy, and basic research into extracellular matrix biology.

Summary Table

Conclusion

• Clone G11-6 is notable for its cross-species reactivity with feline PD-L1 and its utility in detecting PD-L1 expression in specific feline cancers, despite limitations in western blotting due to epitope conformation.
• Clone scFv(G11) stands out for its high-affinity binding to tenascin-C, slow off-rate, and biochemical stability, making it a promising candidate for further therapeutic and diagnostic development.