Anti-Mouse ICOSL (CD275) (Clone HK5.3) – Purified in vivo PLATINUM™ Functional Grade
Anti-Mouse ICOSL (CD275) (Clone HK5.3) – Purified in vivo PLATINUM™ Functional Grade
Product No.: C2448
Clone HK5.3 Target B7-H2 Formats AvailableView All Product Type Monoclonal Antibody Alternate Names ICOSLG, ICOS-L, B7RP-1, LICOS, B7 homolog 2, B7-H2, B7H2, B7h Isotype Rat IgG2a Applications B , FC , in vivo , N |
Antibody DetailsProduct DetailsReactive Species Mouse Host Species Rat Recommended Isotype Controls Recommended Dilution Buffer Immunogen Transfected Cell Line for Mouse B7H2 Product Concentration ≥ 5.0 mg/ml Endotoxin Level <0.5 EU/mg as determined by the LAL method Purity ≥98% monomer by analytical SEC ⋅ >95% by SDS Page Formulation This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration. Product Preparation Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates. Storage and Handling Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles. Country of Origin USA Shipping Next Day 2-8°C RRIDAB_2829462 Applications and Recommended Usage? Quality Tested by Leinco Flow Cytometry For flow cytometric staining, the suggested use of this reagent is ≤1.0 µg per million cells in 100 µL volume or 100 µL of whole blood. Additional Applications Reported In Literature ? Additional reported applications (for the relevant formats) include: Blocking ligand binding Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change. DescriptionDescriptionSpecificity Rat Anti-Mouse ICOSL (CD275) (Clone HK5.3) recognizes an epitope on Mouse CD275. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype. Background ICOSL, also known as CD275 and B7-H2, is a member of the B7 family of co-stimulatory molecules related to B7-1 and B7-2. It is a transmembrane glycoprotein with extracellular IgV and IgC domains. ICOSL is expressed on B cells and macrophages. It binds to ICOS on activated T cells and thus delivers a positive costimulatory signal for optimal T cell function1. ICOS and B7RP-I do not interact with proteins in the CD28-B7 pathway2. The structural features of ICOSL are crucial for its costimulatory function. ICOSL constructs lacking either the IgC or IgV domain demonstrates that receptor binding is mediated solely by the IgV domain but requires the IgC domain for maintaining the structural integrity of the protein3. Antigen Distribution splenic B-cells, T-cells, dendritic cells and macrophages Ligand/Receptor ICOS Function Binds to ICOS. Co-stimulates T cell responses including proliferation and cytokine secretion. Stimulates B cell proliferation and differentiation to plasma cells. PubMed NCBI Gene Bank ID UniProt.org Research Area Immunology Leinco Antibody AdvisorPowered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments. Clone HK5.3 is a monoclonal antibody targeting mouse ICOSL (CD275) and is commonly used in vivo in mice to block the interaction between ICOSL and its receptor ICOS, thereby inhibiting ICOS-mediated co-stimulatory signaling in the immune system. Common in vivo applications in mice include:
In these in vivo settings, HK5.3 is typically administered via intraperitoneal injection or other systemic routes to achieve neutralization/blockade of ICOSL on antigen-presenting cells (e.g., B cells, dendritic cells, macrophages), thereby allowing researchers to dissect the importance of ICOS:ICOSL signaling in various immunological processes. Key details:
No substantial evidence was found for other uses outside these immunological and blocking applications in mice. HK5.3 is widely reported as an in vivo research tool to study the function of ICOSL in mouse models of immunity and disease. Commonly used antibodies and proteins alongside HK5.3 (anti-mouse ICOSL/CD275) in the literature depend on the experimental context. However, studies using HK5.3 generally involve:
Protein partners or related reagents include:
Most co-usage combinations are detailed in immunophenotyping, immune checkpoint, and functional blocking assay publications using the above tools along with HK5.3. If you require a list of specific clone names or additional technical details for standard pairs or experimental controls used with HK5.3, please clarify the research context (e.g., in vivo depletion, flow cytometry, or checkpoint blockade models). The key findings from scientific literature using the HK5.3 clone—a monoclonal antibody specific for mouse ICOSL (CD275/B7-H2)—center on its roles in immunology research, especially regarding T cell costimulation, regulatory T cell homeostasis, and functional blockade studies. Major findings and applications of clone HK5.3:
Summary Table: Key Research Uses of HK5.3
Caveats: Citations found span in vitro and in vivo applications, and effects may depend on model and context (disease state, immune background, etc.). The antibody's functional performance (e.g., required concentration) can vary and is often titrated for each model. Overall, HK5.3 is a well-established research antibody for dissecting the role of ICOSL in mouse immunology, particularly in T cell biology, regulatory T cell maintenance, and immune regulation in health and disease. The dosing regimens for clone HK5.3, an anti-mouse ICOSL (CD275) antibody, vary considerably across different experimental contexts and research objectives in mouse models. Standard Dosing ParametersThe HK5.3 antibody is typically administered via intraperitoneal injection in mouse models. This rat IgG2a monoclonal antibody blocks the binding of ICOSL to ICOS and has been shown to lead to a loss of regulatory T cells in treated mice. Context-Specific Dosing RegimensHydrodynamic Tail Vein (HDTV) Injection Models In studies using HDTV-injected wild-type C57Bl/6 mice, researchers employed a dose of 100 μg per mouse administered intraperitoneally on days 0, 3, 6, and 9 after HDTV injection. This schedule represents a relatively frequent dosing approach with treatment occurring every 2-3 days during the acute experimental phase. Atherosclerosis Models For long-term treatment in apolipoprotein E-deficient mice fed a western diet, a different strategy was used. Mice received α-ICOSL antibody twice a week for 6 weeks. This represents a more sustained, lower-frequency dosing regimen appropriate for chronic disease models where prolonged ICOSL blockade is desired. ADAM10-Mediated Shedding Studies When investigating ICOSL shedding on B cells, researchers utilized a sub-optimal dose of 10 μg per mouse administered every other day. This notably lower dose compared to the 100 μg standard was deliberately chosen to reduce exposure levels while maintaining sufficient biological activity to observe experimental effects. Comparison to Related AntibodiesThe dosing of HK5.3 differs from other immune checkpoint and depletion antibodies commonly used in mouse models. For instance, anti-CTLA-4 antibodies typically use 100-250 μg per mouse every 3 days, while CD4 or CD8 depletion antibodies require 200-250 μg per mouse administered 2-3 times per week. The variability in HK5.3 dosing—from 10 μg to 100 μg—reflects the different biological endpoints being measured, whether complete ICOSL blockade or partial modulation of the ICOS-ICOSL pathway is required. References & Citations1. Wiendl, H. et al. (2003) Brain 126:1026 2. Yoshinaga, SK. et al. (1999) Nature 402:827 3. Chattopadhyay, KJ. et al. (2006) Immunol. 177:3920 Technical ProtocolsCertificate of Analysis |
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