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
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.
Description
Description
Specificity
MIG-2F5-5 activity is directed against murine CXCL9 (monokine induced by gamma interferon, MIG).
Background
CXCL9 is a chemokine, which are small 8-15 kDa proteins that function in immune responses1. CXCL9, -10, -11 and their receptor CXCR3 regulate immune cell migration, differentiation, and activation, leading to tumor suppression in the paracrine axis. However, in the autocrine axis, they may be involved in tumor growth and metastasis. The CXCL9, -10, -11/CXCR3 axis also regulates differentiation of naïve T cells to T helper 1 (Th1) cells. CXCL9, -10, and -11 are usually expressed at low levels but are upregulated by cytokine stimulation. CXCL9 is dependent on IFNγ for expression2. CXCL9 is also capable of direct antimicrobial activity against pathogen infection3.
CXCL9 is secreted by macrophages4, monocytes, endothelial cells, fibroblasts, and cancer cells in response to IFN-γ1 and is also expressed in intratumoral dendritic cells5. CXCL9 is also detectable in CD103+ conventional dendritic cells (cDCs) isolated from transgenic murine MMTV-PyMT tumors following in vivo administration of brefeldin A5. Additionally, CXCL9 is detectable in myeloid cells following ex vivo stimulation with IFN-γ. Furthermore, CXCL9 expression is enhanced in CD8α+ cDC1s when anti-TIM-3 is added. Neutralizing antibodies against Galectin-9 lead to an increase in CXCL9 expression comparable to that induced by anti-TIM-3 antibody. Additionally, endothelial cell expression of CXCL9 is strongly increased in liver sinusoidal endothelial cells isolated from nonalcoholic steatohepatitis mouse livers6.
MIG-2F5-5 was generated by immunizing male Armenian hamsters with recombinant murine CXCL9, and specificity was confirmed by ELISA7.
Antigen Distribution
CXCL9 is mainly secreted by macrophages, monocytes, endothelial cells, fibroblasts, and cancer cells in response to IFN-γ and is also expressed in intratumoral dendritic cells.
Powered 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.
The MIG-2F5-5 clone is an anti-mouse CXCL9 monoclonal antibody commonly used in in vivo studies with mice. Here are some of its common applications:
CXCL9 Neutralization: This antibody is used to neutralize CXCL9, a chemokine crucial for the recruitment of T cells and other immune cells. Neutralizing CXCL9 can help study its role in immune responses and diseases, such as cancer and autoimmune disorders.
Cancer Research: MIG-2F5-5 is applied in cancer models to study the impact of CXCL9 on tumor growth and metastasis. By blocking CXCL9, researchers can assess its role in T cell recruitment to tumors and its potential as a therapeutic target.
Immunotherapy Studies: The antibody helps investigate the effects of immunotherapies, such as those involving checkpoint inhibitors or cytokine therapies, by modulating the CXCL9-CXCR3 axis. This axis is important for T cell trafficking and activation in the context of tumors.
Autoimmune Disease Models: In autoimmune diseases, MIG-2F5-5 can be used to study the role of CXCL9 in inflammation and immune cell infiltration into tissues, helping to understand disease mechanisms and potential treatments.
Vaccine Development: Researchers may use this antibody to assess how CXCL9 influences vaccine efficacy by modulating immune cell trafficking and activation in response to vaccine antigens.
MIG-2F5-5 (anti-mouse CXCL9) is most commonly used in combination with antibodies and proteins that characterize the CXCL9/CXCR3 chemokine axis and related immune pathways. Key examples include:
CXCL10 and CXCL11: These are other chemokines in the same functional group as CXCL9 and also bind to the CXCR3 receptor. Measuring these together helps dissect the entire chemokine axis involved in T-cell recruitment and inflammation.
CXCR3: The primary receptor for CXCL9, CXCL10, and CXCL11. Antibodies to CXCR3 are frequently used to identify and characterize the responsive cell populations, especially activated and memory CD4+ and CD8+ T cells and NK cells.
IFN-γ (Interferon gamma): This cytokine is a key inducer of CXCL9 expression and is often measured to establish the upstream regulation of the chemokine response.
Immune checkpoint molecules: Antibodies against molecules such as TIM-3 and Galectin-9 are commonly used in studies examining immune exhaustion and regulation, especially in the context of chronic infection, cancer, or inflammation.
Applications and additional markers:
MIG-2F5-5 is often used in intracellular flow cytometry together with these and other markers to map immune cell activation, chemotactic responses, and inflammatory states.
Related markers in panel design may include general T cell, memory, or activation markers (e.g., CD4, CD8, CD44, CD69), and markers distinguishing various myeloid or dendritic cell populations that produce or respond to CXCL9.
In summary, other commonly used antibodies or proteins in the literature with MIG-2F5-5 include CXCL10, CXCL11, CXCR3, IFN-γ, TIM-3, and Galectin-9, as well as general T and NK cell markers depending on the immune context.
Key scientific findings from citations using clone MIG-2F5-5 highlight its critical role in the study of the chemokine CXCL9 (MIG), particularly in immunology and cancer research.
Cancer Immunotherapy: Studies using clone MIG-2F5-5 have demonstrated that CXCL9 expression, regulated via the IFN-I pathway, is strongly correlated with T cell expansion and improved clinical outcomes following PD-(L)1 checkpoint blockade in human cancer patients. This underscores CXCL9 as an important biomarker and mechanistic player in effective antitumor immune responses.
Chemotaxis and T Cell Recruitment: MIG-2F5-5 is frequently used to show that CXCL9 acts as a chemoattractant for T cells and NK cells expressing its receptor, CXCR3, and is involved in guiding these effector immune cells into inflamed tissues and tumors. This function is fundamental to inflammatory responses and tumor immune infiltration.
Functional Studies and Neutralization: The clone is validated for a variety of applications, including intracellular staining, flow cytometry, in vivo neutralization, and immunofluorescence, allowing researchers to profile and functionally dissect CXCL9’s role in diverse biological contexts such as tumor microenvironment, infection, and inflammatory diseases.
Cellular Sources and Regulation: Citation-supported research using this antibody has mapped CXCL9 production primarily to monocytes, macrophages, endothelial cells, fibroblasts, and cancer cells, particularly under IFN-γ stimulation. Sources also highlight that tumor-intrinsic factors (such as CCL5) drive CXCL9 expression in tumor-associated macrophages, influencing immune phenotypes and therapy response.
Broader Phenotypic Effects: Beyond chemotaxis, CXCL9 has roles in T-cell activation, angiogenesis inhibition, and modulation of the immune landscape in both health and disease, as established in studies using MIG-2F5-5.
Technical and Reagent Details: The clone is consistently described as Armenian hamster IgG with high specificity for murine CXCL9, reliable performance in in vivo and in vitro assays, and low endotoxin content, making it ideal for sensitive functional studies.
Summary: Clone MIG-2F5-5 is a validated and widely cited tool antibody pivotal in establishing the immunological significance of CXCL9, particularly in T-cell chemotaxis, antitumor immunity, and biomarker discovery for immunotherapy responsiveness.
Dosing Regimens of Clone MIG-2F5-5 in Mouse Models
Variability in Dosing There is no universal dosing regimen for the anti-mouse CXCL9 (MIG-2F5-5) monoclonal antibody across all mouse models—dosing must be customized based on experimental design, disease model, and published protocols. This reflects the general best practice for in vivo antibody studies, where optimal dosing is often determined empirically and may differ depending on factors such as antibody pharmacokinetics, target engagement, and the biological context of the model.
Published Dosing Example One specific example from the literature uses the MIG-2F5.5 clone at 200 µg/mouse, administered every 3 days for six doses, in a model investigating the impact of CXCL9 neutralization on immune responses. This regimen was chosen to maintain sufficient antibody levels for CXCL9 neutralization throughout the experimental timeline. However, this protocol may not be directly transferable to other disease models or experimental endpoints without adaptation.
Key Considerations for Dosing When establishing a dosing regimen for MIG-2F5-5 (or similar antibodies) in different mouse models, consider the following:
Antibody Half-Life: The dosing interval should account for the antibody’s half-life in circulation, which can vary based on the antibody’s formulation and the host species.
Model-Specific Factors: The disease model (e.g., tumor, infection, autoimmunity), route of administration, and timing relative to disease induction all influence the optimal dose and schedule.
Literature and Supplier Guidance: Always consult supplier datasheets and published studies for initial dosing recommendations, then adjust based on pilot experiments and observed biological effects.
Endotoxin Levels: For in vivo use, ensure the antibody is of high purity and low endotoxin, as this can affect tolerability and immune responses.
No Standardization Across Labs The lack of standardized dosing reflects the broader reality in preclinical research: antibody dosing is often established on a lab-by-lab, model-by-model basis. Researchers must validate their chosen regimen through pilot experiments, monitoring both efficacy (e.g., target neutralization, disease outcome) and safety (e.g., absence of toxicity, normal behavior).
Summary Table: Example Dosing Regimen
Model Context
Dose per Mouse
Frequency
Total Doses
Reference
Immune response study
200 µg
Every 3 days
6
Recommendations
Start with published regimens as a benchmark, then tailor based on your model’s needs.
Pilot testing is essential to determine the minimal effective dose and optimal scheduling.
Document and justify your dosing rationale in publications and protocols to aid reproducibility.
In summary, dosing regimens for clone MIG-2F5-5 vary significantly across mouse models and must be empirically determined for each experimental context. Published examples exist, but adaptation to specific research questions is necessary.
References & Citations
1. Tokunaga R, Zhang W, Naseem M, et al. Cancer Treat Rev. 63:40-47. 2018.
2. Cole KE, Strick CA, Paradis TJ, et al. J Exp Med. 187: 2009–2021. 1998.
3. Reid-Yu SA, Tuinema BR, Small CN, et al. PLoS Pathog. 11(2):e1004648. 2015.
4. Marcovecchio PM, Thomas G, Salek-Ardakani S. J Immunother Cancer. 9(2):e002045. 2021.
5. de Mingo Pulido Á, Gardner A, Hiebler S, et al. Cancer Cell. 33(1):60-74.e6. 2018.
6. Xiong X, Kuang H, Ansari S, et al. Mol Cell. 75(3):644-660.e5. 2019.
7. Krug A, Uppaluri R, Facchetti F, et al. J Immunol. 169(11):6079-6083. 2002.
8. Asai A, Tsuda Y, Kobayashi M, et al. Infect Immun. 78(10):4311-4319. 2010.
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
