Anti-Mouse CD209b [Clone 22D1] — Purified in vivo PLATINUM™ Functional Grade

The Clone 22D1 is used in in vivo mouse studies primarily to target CD209b (SIGN-R1), a type II transmembrane C-type lectin receptor involved in the innate immune response. This receptor is expressed on specific macrophages, such as those in splenic marginal zones and lymph node medulla, and plays a role in recognizing high-mannose-containing glycoproteins on microbial surfaces. The 22D1 monoclonal antibody is used to block or study CD209b in vivo, as it can bind to and block this receptor's function.

Applications in In Vivo Mouse Studies:

• In Vivo SIGN-R1 Blockade: The antibody is used to block the CD209b receptor in vivo, which can help in studying its role in immune responses, such as the uptake of pathogens by macrophages.
• Immunohistochemistry and Western Blot: The antibody is applicable for histological studies and protein analysis in tissues.
• Flow Cytometry: It can be used to analyze the expression of CD209b on specific cell populations in mice.

Overall, the Clone 22D1 is a valuable tool for researching the immune system and its components, particularly in the context of CD209b and its role in mouse models.

Based on the search results, the sterile packaged clone 22D1 (Anti-mouse CD209b antibody) should be stored at -20 to -70°C for optimal shelf life.

Storage Temperature Guidelines

The clone 22D1 antibody has specific temperature requirements that differ from general medication storage guidelines. While most medicines should be stored at 59 to 77 degrees F (15 to 25°C) at room temperature, biological products like antibodies require much colder storage conditions to maintain their stability and effectiveness.

Recommended Storage Conditions

For the clone 22D1 antibody, the storage recommendations are:

• Long-term storage: -20 to -70°C as supplied, which provides 12 months of stability from the date of receipt
• Short-term storage: 2 to 8°C (refrigerated), which allows for 1 month of stability from the date of receipt

Important Storage Considerations

When storing this biological product, it's crucial to use a manual defrost freezer and avoid repeated freeze-thaw cycles, as these can damage the antibody's structure and reduce its effectiveness. Unlike general medications that can tolerate room temperature storage, antibodies and other biological products require consistent cold storage to maintain their molecular integrity and biological activity.

The sterile packaging helps protect the product from contamination, but the temperature control remains the most critical factor for preserving the antibody's functionality over time.

Commonly Used Antibodies and Proteins Alongside 22D1 in Research

The "22D1" clone refers to a monoclonal antibody (mAb) specifically targeting mouse CD209b (also known as SIGN-R1), a type II transmembrane C-type lectin expressed in specific macrophage populations. The most commonly cited use of 22D1 in the literature is in flow cytometry (FACS) for the detection and study of splenic marginal zone macrophages and lymph node medullary macrophages. In these contexts, several other antibodies and proteins are frequently used in combination with 22D1, primarily to characterize immune cell subsets or investigate functional pathways.

Co-staining Antibodies for Immune Cell Characterization

When using 22D1 for immunofluorescence or flow cytometry, researchers typically co-stain with antibodies against other cell surface markers to identify and isolate specific immune cell populations. While the specific combinations are not detailed in the provided sources, general practice in immunology (especially for macrophages and marginal zone cells) includes:

• Anti-F4/80 and anti-CD11b: For general macrophage identification.
• Anti-CD19 and anti-B220: For B cell identification.
• Anti-CD3 and anti-TCR: For T cell identification.
• Anti-MHC II (I-A/I-E): For antigen-presenting cell identification.
• Anti-CD11c: For dendritic cells.

These are not explicitly linked to 22D1 in the provided sources, but given the expression profile of CD209b, such co-staining would be standard in the literature for comprehensive immunophenotyping.

Functional Pathway Proteins Associated with SIGN-R1 (CD209b)

The 22D1 antibody is used to study not only cellular identity but also functional interactions, particularly concerning SIGN-R1’s role in pathogen recognition and complement activation. The following proteins are directly cited as interacting partners or pathway members:

• TLR4/MD2: SIGN-R1 (CD209b) physically associates with TLR4/MD2, implicating it in the recognition of bacterial lipopolysaccharide (LPS).
• Complement C1q and C3: SIGN-R1 binds directly to the complement component C1q to assemble a non-conventional C3 convertase, affecting complement activation and catabolism.
• Dextran polysaccharides and Streptococcus pneumoniae capsular polysaccharide: These are ligands directly recognized and internalized by SIGN-R1.

These protein interactions are often studied using 22D1 in flow cytometry, Western blot, or co-immunoprecipitation experiments to delineate SIGN-R1’s role in innate immunity.

Technical and Control Antibodies

When performing assays with 22D1, the following are commonly used as controls or for experimental validation:

• Isotype controls: Armenian hamster IgG is the isotype for 22D1, and matching isotype control antibodies are essential for flow cytometry and immunohistochemistry.
• Secondary detection reagents: For example, anti-Armenian hamster IgG secondary antibodies for detection in IHC or IF, although 22D1 is also available as a direct conjugate (e.g., Alexa Fluor 488).
• Loading controls: For Western blotting (e.g., beta-actin, GAPDH), though not specific to 22D1, these are standard in any protein detection workflow.

Summary Table of Common Partners

Conclusion

The 22D1 antibody is primarily used in the context of mouse splenic and lymph node macrophage research, often in combination with antibodies against general macrophage, B cell, and T cell markers for immunophenotyping. Functionally, it is studied alongside proteins involved in pathogen recognition (TLR4/MD2), complement activation (C1q, C3), and specific polysaccharide ligands. Proper controls, including isotype-matched antibodies and appropriate secondary reagents, are standard in these experiments.

Based on the scientific literature, there are two distinct research areas involving "clone 22" terminology that have yielded significant findings:

TSC22D1 Family Proteins Research

The most substantial findings relate to TSC22D1 (TGF-? stimulated clone-22) family proteins, which are important regulators of cellular processes. TSC-22 has been reported to induce differentiation, growth inhibition, and apoptosis in various cells. This protein family consists of multiple variants produced from four different family genes, with TSC-22 (TSC22D1-2) being composed of 144 amino acids translated from a short variant mRNA of the TSC22D1 gene.

Subcellular Localization Patterns

Research has revealed distinct intracellular distribution patterns for different TSC22D1 family members. TSC22D1-1 was found to be predominantly localized in the nucleus, while TSC-22 (TSC22D1-2) was primarily localized in the cytoplasm, particularly in mitochondria, and notably translocated from the cytoplasm to the nucleus following DNA damage. TSC22(86) (TSC22D1-3) demonstrated dual localization in both cytoplasm and nucleus.

Protein-Protein Interactions

Mass spectrometry analyses have identified multiple binding protein candidates for TSC22D1 family proteins through both in vitro pull-down assays and in vivo binding assays. Two key interactions have been confirmed:

• Histone H1 binds to TSC-22 (TSC22D1-2) or TSC22(86) (TSC22D1-3) in the nucleus
• Guanine nucleotide-binding protein-like 3 (GNL3), also known as nucleostemin, binds to TSC-22 (TSC22D1-2) in the nucleus

Proposed Mechanisms

The research suggests potential mechanisms whereby TSC22D1 family proteins may promote histone H1 degradation through their interaction, with replacement of chromatin binding by HMGA2 potentially causing changes in gene expression that regulate cell proliferation, differentiation, apoptosis induction, and cell senescence. Additionally, GNL3's interaction with TSC-22 may influence cell cycle progression and cellular differentiation.

Code Clone Detection Research

In a separate field of computer science, research on code clone detection using recurrent neural networks has shown that RNN techniques demonstrate effectiveness in detecting challenging clone types. Recent studies have shown enhanced performance in identifying Type-III and Type-IV clones, with improvements particularly evident in precision, recall, and F1-scores. Among the techniques evaluated, the DFS technique demonstrated the highest F1-score performance in detecting Type-III and Type-IV clones, surpassing peers in both BigCloneBench and OJClone benchmarks.

The findings indicate notable improvement in RNN-based clone detection performance over time, with relatively lower F-scores in 2017 and 2018 attributed to the early developmental stage of deep learning techniques in this domain.