Anti-Mouse CD317 [Clone 927] — Purified in vivo GOLD™ Functional Grade

Use of Clone 927 in In Vivo Mouse Studies

Clone 927 is a rat IgG2b, ? monoclonal antibody specific for murine CD317 (also known as BST2 or PDCA-1), a type II transmembrane glycoprotein. Its primary application in in vivo mouse studies is for the specific depletion of plasmacytoid dendritic cells (pDCs), which are the main cell type expressing CD317 in naïve mice. Here’s how clone 927 is utilized:

Mechanism and Applications

• Specific pDC Depletion: When administered in vivo, clone 927 effectively depletes mouse plasmacytoid dendritic cells by targeting the CD317 antigen, which is expressed almost exclusively on these cells in unstimulated animals.
• Functional Outcomes: In vivo treatment with mAb 927 (clone 927) leads to ~95% depletion of pDCs and significantly reduces IFN-? secretion by these cells upon stimulation with CpG, demonstrating the critical role of pDCs in type I interferon production.
• Experimental Utility: The mouse CD317 antigen can also be expressed on some tumor cells and is inducible on a variety of other cell types under certain conditions, but in naïve mice, it serves as a selective marker for pDCs. This makes clone 927 a powerful tool for interrogating the role of pDCs in immune responses, infection, cancer, and autoimmunity.

Technical Details

• Isotype: Rat IgG2b, ?
• Formulation: Generally supplied in a low endotoxin, purified form suitable for in vivo injection.
• Recommended Use: Clone 927 is used for in vivo depletion studies, immunofluorescence, and flow cytometry; its functional grade ensures minimal off-target effects and reliable depletion.
• Control: An appropriate isotype control (rat IgG2b, anti-keyhole limpet hemocyanin) is recommended for comparison.

Summary Table

Conclusion

Clone 927 is a well-validated, highly specific tool for depleting plasmacytoid dendritic cells in mice in vivo, enabling researchers to dissect the role of these cells in immunity and disease with minimal perturbation to other cell types. Its use is standard in studies requiring selective pDC ablation to understand their contributions to antiviral responses, tumor immunology, and autoimmune processes.

Based on the available information, clone 927 is an anti-mouse CD317 monoclonal antibody that recognizes CD317 (also known as BST2, PDCA-1, or tetherin) on plasmacytoid dendritic cells and other cell types. However, the search results do not provide specific information about other antibodies or proteins commonly used in combination with the 927 antibody in research applications.

Primary Applications of Clone 927

The 927 antibody is primarily used for studying interferon-producing cells (IPCs), also known as plasmacytoid dendritic cells, which are early responders to viral infection. The antibody has been validated for several applications including flow cytometry, immunofluorescence microscopy, functional assays, and in vivo depletion studies.

Target Expression and Function

CD317, the target antigen of clone 927, is expressed on various cell types including T cells, mast cells, B cells, fibroblast cells, and pluripotent embryonal carcinoma cells. The protein functions as an antiviral factor that blocks the release of nascent retrovirus particles from infected cells. When mice are challenged with stimuli such as influenza, CpG, LPS, or other immune activators, CD317 expression extends to dendritic cells, myeloid cells, T cells, B cells, NKT cells, and some NK cells.

Potential Research Context

While specific combinations aren't detailed in the search results, the 927 antibody would likely be used alongside other markers for comprehensive immunophenotyping studies. Given that it targets plasmacytoid dendritic cells, researchers might combine it with other dendritic cell markers, B cell markers (since it's also expressed on CD138+ plasma cells), or interferon pathway-related proteins to study immune responses to viral infections or inflammatory conditions.

The information available does not provide a comprehensive list of commonly co-used antibodies or proteins with the 927 clone in published literature.

Key Findings from Clone 927 Scientific Citations

Clone 927 most prominently refers to a monoclonal antibody (anti-mouse CD317, Clone 927) used primarily in immunology research. The scientific literature and product documentation highlight several critical findings and applications related to this clone:

Primary Applications and Biological Insights

• Target Specificity: Clone 927 specifically recognizes murine CD317 (also known as BST2, tetherin, HM1.2 antigen, bone marrow stromal antigen 2, or PDCA-1). CD317 is a key marker for interferon-producing cells (IPCs), which are a subset of plasmacytoid dendritic cells (pDCs) crucial in early antiviral immune responses.
• Identification of Rare Immune Cells: The antibody was developed to overcome challenges in identifying and studying IPCs, which are rare in blood and tissues and have a complex surface phenotype. Clone 927 enables precise detection and isolation of these cells for further study.
• Functional Effects In Vivo: Administration of mAb 927 in vivo leads to significant effects on the immune system: it abrogates IFN-? secretion by IPCs in response to CpG stimulation and depletes IPCs by approximately 95%, markedly reducing plasma cells. This makes it a valuable tool for studying the role of IPCs/pDCs in immune responses and viral infections.
• Cell Surface and Intracellular Localization: CD317 is expressed on the cell surface of murine IPCs under naive conditions and is also found intracellularly in the Golgi apparatus, suggesting a role in protein sorting between the Golgi and plasma membrane.
• Broad Expression Pattern: While primarily a marker of IPCs in naive mice, CD317 expression can be induced on most cell types by stimulation with type I interferons and IFN-?.

Technical and Experimental Details

• Antibody Generation: Clone 927 was generated by immunizing rats with bone marrow-derived IPCs, followed by hybridoma creation and screening for specificity to CD11c?B220?Ly-6c?CD11b? splenocytes.
• Isotype and Applications: The antibody is of rat IgG2b isotype and is suitable for various applications including flow cytometry, immunofluorescence, immunohistochemistry, and in vivo depletion studies.

Summary Table

Conclusion

Clone 927 is a critical reagent for the study of murine plasmacytoid dendritic cells and interferon responses, enabling researchers to identify, isolate, and functionally manipulate these rare but important immune cells. Its development has addressed significant technical barriers in IPC research, providing a robust tool for both basic and translational immunology studies.

Clone 927 (anti-mouse CD317, also known as BST2 or PDCA-1) is primarily used for in vivo depletion of plasmacytoid dendritic cells (pDCs) in mouse models, but published sources do not specify a universally standardized dosing regimen; instead, dose and schedule can vary by experimental context and mouse strain.

Key context and dosing details:

• Mechanism: Clone 927 depletes pDCs (~95%) and abrogates IFN-? secretion by these cells in response to immune stimuli such as CpG.
• Typical application: Used in tumor immunology, infection, and autoimmunity studies to study the role of pDCs.
• Reported usage: Specific dosing regimens for clone 927 are not widely detailed on reagent supplier or antibody product pages. Product technical sheets and research summaries confirm its use for in vivo depletion but omit standard dose and frequency parameters.

Dosing regimens in related antibody models:

To infer typical practices for clone 927, comparison with similar in vivo depletion antibodies in murine models provides guidance:

• Depleting antibodies (e.g., GK1.5 for CD4, 2.43 for CD8, RB6-8C5 for Gr-1) are most frequently dosed at 200–250 ?g per mouse, intraperitoneally (IP), 2–3 times per week, depending on the desired degree and duration of depletion.
• These regimens are used across various mouse models (including C57BL/6, BALB/c, and tumor-bearing mice).

Variation by mouse model:

• Most depletion antibodies (including those for pDCs) are dosed similarly across common laboratory strains unless specific differences in mouse size, immune status, or experimental protocol require adjustment.
• There is no evidence of significant PK/PD differences in antibody-based depletion when adjusting standard dosing regimens between mouse models in published comparisons.

Recommendations:

• In the absence of a published, universally accepted dosing protocol for clone 927, a starting regimen of 200–250 ?g per mouse, injected IP, 2–3 times per week aligns with best practice for similar depleting antibodies.
• Dosing may be adjusted depending on the mouse strain (e.g., smaller strains may warrant the lower end of the dosing range) and study-specific goals (e.g., acute versus chronic depletion).
• Always consult pilot depletion/efficacy data and review relevant recent literature if using unique models (e.g., immunodeficient, tumor-bearing, or genetically modified mice) to optimize the schedule.

Summary Table: Standard Depletion Antibody Regimens (for reference):

If a unique strain or disease model is being used, consult primary literature or pilot titration experiments to confirm full depletion and minimize off-target effects.

Direct experimental guidance for clone 927, such as peer-reviewed publications or protocol-specific recommendations, may provide further refinement; absence of such data warrants careful, measured empirical optimization.