Anti-Mouse CD276 (Clone MJ18) – Purified in vivo GOLD™ Functional Grade
Anti-Mouse CD276 (Clone MJ18) – Purified in vivo GOLD™ Functional Grade
Product No.: C3346
Clone MJ18 Target CD276 Formats AvailableView All Product Type Monoclonal Antibody Alternate Names CD276; CD276 antigen; B7h3; B7RP-2; AU016588; 6030411F23Rik Isotype Rat IgG1 κ Applications B , FC , in vivo |
Antibody DetailsProduct DetailsReactive Species Mouse Host Species Rat Recommended Isotype Controls Recommended Dilution Buffer Immunogen Mouse B7-H3 IgG2a fusion protein Product Concentration ≥ 5.0 mg/ml Endotoxin Level <1.0 EU/µg as determined by the LAL method Purity ≥95% 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 Applications and Recommended Usage? Quality Tested by Leinco FC Additional Applications Reported In Literature ? B FC Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change. DescriptionDescriptionSpecificity MJ18 activity is directed against mouse CD276 also known as B7-H3. Background CD276, also known as B7 homolog 3 protein (B7-H3), is a member of the B7 superfamily and acts as an immune checkpoint molecule and a costimulatory/coinhibitory immunoregulatory protein1. CD276 influences innate and adaptive immunity, regulates the aggressiveness of cancer cells, and is thought to play an important role in tumor development and cancer immunity. CD276 has been studied in many cancers, including breast, lung, ovarian, brain, gastric, and squamous cell carcinoma.
Human CD276 exists as either a soluble isoform or as a ~45–66 kDa type I transmembrane protein that is composed of an extracellular domain, a transmembrane domain, and a short intracellular domain1. In murine CD276, the extracellular domain is composed of a single pair of immunoglobulin variable and constant domains. Soluble CD276 is produced by cleavage from the cell surface or via alternative intron splicing and has been found in the secretomes of exosomes and other extracellular vesicles. In normal human tissues, CD276 mRNA is widely and abundantly expressed but protein abundance is low1. miR-124 is thought to cause translational repression of CD276 by targeting its 3’-UTR, while other miRNAs are known to affect CD276 expression. In contrast, in tumor cells, CD276 mRNA and protein are abundant, and its presence is correlated with worsened prognosis, poor survival, recurrence rate, and enhanced invasive and migratory properties. CD276 blocking with monoclonal antibodies has been shown to reduce tumor growth and prolong survival in mouse models of various cancers. TREM-like transcript 2 (TLT-2) has been identified as a potential receptor2. MJ18 was generated by immunizing Sprague Dawley rats with aa 1–242 of mouse CD276 extracellular domain linked to the Fc portion of mouse IgG2a3. After immunization, lymph node cells were fused with P3U1 myeloma cells, hybridomas were selected by flow cytometry, and MJ18 was purified from ascites. Antigen Distribution CD276 is weakly expressed on activated lymphocytes, macrophages, dendritic cells, nasal and airway epithelial cells, osteoblasts, and some tumor cell lines. A soluble form is secreted by monocytes, dendritic cells, and activated T cells. 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 MJ18 is commonly used in vivo in mice for applications aimed at blocking the function of CD276 (B7-H3), an immune checkpoint molecule thought to dampen T cell activity and promote tumor immune evasion. The intended uses have included:
Critical context:
Summary of reported in vivo uses:
Important limitation: In the scientific literature, several antibodies and proteins are commonly used alongside MJ18. These include:
These proteins and antibodies are utilized in various research contexts, including immunological studies and cancer research, to better understand immune responses and potential therapeutic targets. The key findings from scientific literature regarding clone MJ18 are that MJ18, although widely cited as a blocking antibody for murine B7-H3 (CD276), actually does not bind B7-H3, but instead recognizes the Fc receptor FcγRIIB on murine splenocytes, predominantly on B cells. Essential context and supporting details:
Additional relevant information:
In summary, MJ18 does not bind B7-H3 and instead targets FcγRIIB, underscoring a significant correction to past scientific practice and literature regarding its use. Based on available evidence, dosing regimens of clone MJ18 show minimal variation across different mouse models, with researchers typically applying a standardized protocol regardless of the specific experimental context. Standard Dosing ProtocolThe most commonly reported dosing regimen for MJ18 is remarkably consistent: 300 μg per injection administered every other day via intraperitoneal injection. This protocol has been adopted as the standard approach across various experimental settings, including genetically modified mouse models and syngeneic tumor systems. For example, in studies using Tgfbr1/Pten 2cKO mice (a genetically engineered model with mixed FVBN/CD1/129/C57 background), researchers administered 0.3 mg of MJ18 intraperitoneally every other day starting from day 14 after tamoxifen induction. Similarly, in orthotopic rhabdomyosarcoma models (M3-9-M), investigators used the identical 300 μg per injection every other day regimen. Lack of Model-Specific VariationsNo significant published evidence suggests major variations of the MJ18 dosing regimen based on different mouse strains or tumor models. This uniformity persists despite differences in:
The standardized approach contrasts with the model-specific optimization often seen with other checkpoint blockade antibodies, where doses may range significantly (for example, anti-PD-1 antibodies typically use 100-500 μg per mouse depending on the clone and application). Important CaveatRecent research has revealed that MJ18 does not actually recognize B7-H3 on the surface of mouse cells, despite being widely used as an anti-B7-H3 blocking antibody. Instead, MJ18 appears to bind Fc receptors on immune cells, particularly B cells. This finding raises questions about the mechanistic basis for any observed effects and suggests that the uniform dosing regimen may reflect convention rather than optimized targeting of a specific antigen. References & Citations1. Zhou WT, Jin WL. Front Immunol. 12:701006. 2021.
2. Hashiguchi M, Kobori H, Ritprajak P, et al. Proc Natl Acad Sci USA. 105: 10495–10500. 2008. 3. Nagashima O, Harada N, Usui Y, et al. J Immunol. 181(6):4062-71. 2008. 4. Yamato I, Sho M, Nomi T, et al. Br J Cancer. 101(10):1709-1716. 2009. 5. Kamachi F, Isshiki T, Harada N, et al. Biochem Biophys Res Commun. 463(4):739-45. 2015. Technical ProtocolsCertificate of Analysis |
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