LGALS2 Mouse, Active
Description
Role in Colorectal Cancer and Colitis
LGALS2 exhibits dual roles in colorectal disease:
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Colitis Suppression: Gal2-deficient (Gal2-KO) mice showed reduced disease activity index (DAI), lower mortality, and less histological damage in dextran sodium sulfate (DSS)-induced colitis . Exogenous Gal2 treatment has also been reported to induce T-cell apoptosis, alleviating inflammation .
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Tumor Promotion: In the azoxymethane (AOM)/DSS model, Gal2-KO mice developed larger tumors with elevated STAT3 phosphorylation, suggesting Gal2 suppresses tumor growth via STAT3 inhibition .
Involvement in Arteriogenesis and Macrophage Polarization
LGALS2 regulates vascular remodeling and immune cell behavior:
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Collateral Artery Growth: Anti-Gal2 antibodies (e.g., 2H8, 2C10) enhanced arteriogenesis in LDLR-null mice by promoting M2-like macrophage polarization, increasing perivascular M2 fractions from 49% to 73–75% .
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Immune Modulation: Gal2 inhibits M1 macrophage cytokines (e.g., TNF-α, IL-6) and promotes M2 markers (e.g., Arg1, Mgl1) .
Implications in Breast Cancer and Immunotherapy
LGALS2 drives immunosuppression in triple-negative breast cancer (TNBC):
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Tumor Microenvironment (TME): Overexpression of LGALS2 in 4T1/EMT6 cells increased tumor-associated macrophages (TAMs), reduced CD8+ T-cell cytotoxicity, and promoted M2 polarization via CSF1/CSF1R signaling .
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Therapeutic Targeting: Anti-LGALS2 antibodies reversed immune suppression, reducing tumor volume by 50% and increasing granzyme B+/IFN-γ+ T cells .
Therapeutic Potential and Challenges
Product Specs
Galectin-2, also known as LGALS2, belongs to the galectin family. These proteins are defined by their ability to bind carbohydrates (lectins) and play roles in cell-to-cell adhesion, cell-to-extracellular matrix interactions, pre-mRNA splicing, apoptosis, and tumor progression. Galectin-2 specifically has been shown to induce apoptosis in activated T cells, bind to lymphotoxin-a, and may be implicated in myocardial infarction. Human and mouse LGALS2 share approximately 65% amino acid sequence similarity.
Recombinant Mouse LGALS2, expressed in E. coli, is a single, non-glycosylated polypeptide chain. This protein contains 153 amino acids (including a 23 amino acid His-tag at the N-terminus, spanning residues 1-130 of the LGALS2 sequence) and has a molecular weight of 17.3 kDa. The protein is purified using proprietary chromatographic techniques.
LGALS2 protein is supplied at a concentration of 1 mg/ml in a buffer containing 20mM Tris-HCl (pH 8.0), 0.1M NaCl, 1mM DTT, and 10% glycerol.
For short-term storage (2-4 weeks), the product can be stored at 4°C. For long-term storage, it is recommended to store the protein at -20°C. The addition of a carrier protein (0.1% HSA or BSA) is suggested for extended storage. Avoid repeated freeze-thaw cycles.
Purity is determined to be greater than 95% by SDS-PAGE analysis.
The biological activity of LGALS2 is assessed by its ability to agglutinate human red blood cells. The ED50, defined as the concentration required to achieve 50% agglutination, is greater than or equal to 20 µg/ml.
Galectin-2, Gal-2, Lgals2, 2200008F12Rik, AI324147.
MGSSHHHHHH SSGLVPRGSH MGSMSEKFEV KDLNMKPGMS LKIKGKIHND VDRFLINLGQ GKETLNLHFN PRFDESTIVC NTSEGGRWGQ EQRENHMCFS PGSEVKITIT FQDKDFKVTL PDGHQLTFPN RLGHNQLHYL SMGGLQISSF KLE
Q&A
What is LGALS2 and what is its expression pattern in mouse tissues?
LGALS2 (Lectin, Galactoside-Binding, Soluble, 2), commonly known as Galectin-2 or Gal-2, is a soluble beta-galactoside binding lectin that functions as a homodimer. In mice, LGALS2 shows highest expression in the gastrointestinal (GI) tract, particularly in the colon . The protein is involved in multiple biological processes, including modulation of oxidative stress responses and immune cell function. Analysis of mouse tissues reveals that Gal2 expression is tissue-specific, with expression patterns distinct from other galectin family members that are also present in the mouse colon .
What are the known physiological functions of LGALS2 in mouse models?
Studies utilizing Gal2 knockout mouse lines have demonstrated that while LGALS2 is highly expressed in specific tissues, Gal2-KO mice are viable and fertile with no obvious gross abnormalities . This suggests potential functional redundancy with other galectins or context-dependent roles. Experimental evidence indicates LGALS2 plays an important modulatory role in acute colitis and colon tumor development . Additionally, LGALS2 has been identified as part of a ROS-responsive gene network, with its depletion rendering cells more resistant to hydrogen peroxide challenge, indicating involvement in oxidative stress response pathways .
How do researchers distinguish LGALS2 from other galectin family proteins?
Distinguishing LGALS2 from other galectin family members requires:
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Expression analysis: LGALS2 shows preferential expression in the GI tract, while other galectins may have broader expression patterns
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Molecular weight determination: Gal2 has a distinctive molecular weight compared to other family members
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Specific antibody selection: Using antibodies that specifically recognize unique epitopes in LGALS2 rather than conserved regions shared with other galectins
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Knockout validation: Confirming antibody specificity using tissues/cells from LGALS2 knockout mice as negative controls
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Functional assays: Assessing specific binding properties and biological activities characteristic of LGALS2
What techniques are most effective for detecting LGALS2 expression in mouse tissues?
Multiple complementary approaches are recommended for robust LGALS2 detection:
How can researchers generate and validate LGALS2 knockout mouse models?
Generation of LGALS2 knockout mice typically involves:
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Design approach: Target exons 2 and 3 of the LGALS2 gene for deletion, as demonstrated in previous studies
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Validation methods:
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Control considerations: Include both wild-type and heterozygous littermates as controls, as heterozygotes show similar Gal2 expression to wild-type mice
What are the recommended protocols for studying LGALS2 in oxidative stress models?
Based on published research, the following protocols have proven effective:
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In vitro cellular models:
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In vivo approach:
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Use LGALS2 knockout mice with appropriate controls
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Induce oxidative stress through chemical agents or disease models
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Analyze tissue damage, inflammatory markers, and cellular responses
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Compare outcomes between wild-type and knockout animals to determine LGALS2's protective or detrimental effects
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How does LGALS2 influence colorectal cancer development in mouse models?
LGALS2 demonstrates a tumor-suppressive role in colorectal cancer, with several key observations:
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Tumor growth effects: Depletion of LGALS2 promotes colorectal tumor growth in mouse models
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Inflammation-cancer connection: LGALS2 depletion attenuates acute colitis while promoting colorectal tumor growth, suggesting complex context-dependent functions
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Mechanistic insights: The suppressive role of LGALS2 in colon tumor growth highlights its potential as a therapeutic target
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Experimental approach: Studies typically use chemical-induced colon cancer models or xenograft approaches comparing growth rates of tumors with varying LGALS2 expression levels
What is the role of LGALS2 in breast cancer immunology research?
Recent research has identified LGALS2 as a key immunomodulator in breast cancer:
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Immune escape mechanism: LGALS2 has been identified through in vivo CRISPR screens as a candidate regulator in triple-negative breast cancer (TNBC) involving immune escape
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Tumor growth dynamics:
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Immunosuppressive effects: Tumor cell-intrinsic LGALS2 induces increased numbers of tumor-associated macrophages and promotes their M2-like polarization and proliferation through the CSF1/CSF1R axis
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Therapeutic potential: Blockade of LGALS2 using inhibitory antibodies successfully arrests tumor growth and reverses immune suppression in mouse models
How does manipulation of LGALS2 affect the tumor microenvironment in mouse cancer models?
LGALS2 substantially influences the tumor immune microenvironment:
| Immune Cell Population | Effect of LGALS2 Overexpression | Effect of LGALS2 Knockout |
|---|---|---|
| CD8+ T cells (CTLs) | Decreased frequency and cytotoxic function, increased exhaustion | Increased infiltration and activity |
| Natural killer cells | Decreased cytotoxicity | Enhanced anti-tumor function |
| Macrophages | Increased M2-like polarization, upregulated Arg1, Mgl1, and Fizz1 markers | Reduced M2 phenotype |
| Myeloid-derived suppressor cells | Increased infiltration | Decreased presence |
| Regulatory T cells | Enhanced immunosuppressive population | Reduced immunosuppression |
These findings were consistent across different mouse models (4T1 and EMT6 triple-negative breast cancer cell lines) .
What molecular mechanisms mediate LGALS2's effects on immune cells in the tumor microenvironment?
Research has identified several key molecular mechanisms:
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Macrophage polarization: LGALS2 expression in tumor cells stimulates macrophages to exhibit enhanced proliferation and M2-like polarization
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CSF1/CSF1R axis: LGALS2 modulates the CSF1/CSF1R signaling pathway, which is critical for macrophage recruitment, survival, and polarization
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Transcriptional regulation: Transcriptome profiling of cells with different LGALS2 expression levels identified 110 genes upregulated in LGALS2-overexpressing cells and downregulated in LGALS2-KO cells
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Secreted factors: Concentration of serum LGALS2 protein (sLGALS2) correlates with tumor burden, suggesting potential paracrine signaling mechanisms
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Tertiary lymphoid structures: Recent research has identified LGALS2 as a key marker within tertiary lymphoid structures in breast cancer, with a role in dendritic cell stimulation
How do the paradoxical effects of LGALS2 in different cancer types inform research design?
The contrasting roles of LGALS2 across cancer types necessitate careful research design:
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Context-dependent function: LGALS2 acts as a tumor suppressor in colon cancer but promotes immune evasion in breast cancer , requiring researchers to avoid generalizing findings across cancer types
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Temporal dynamics: LGALS2-KO breast cancer cells show increased tumor mass initially followed by dramatic regression , highlighting the importance of longitudinal studies
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In vitro vs. in vivo discrepancies: LGALS2 manipulation shows minimal effects on cell proliferation in vitro but dramatic effects in vivo , emphasizing the critical importance of the tumor microenvironment
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Mechanistic investigation: Research designs should incorporate both tumor cell-intrinsic and immune component analyses to fully understand LGALS2's complex roles
What new therapeutic strategies are emerging from LGALS2 research in mouse models?
Several promising therapeutic approaches have emerged:
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Antibody-based blockade: Single-domain llama-derived therapeutic antibodies against LGALS2 have demonstrated efficacy in preclinical models, significantly reducing tumor volume and weight while enhancing anti-tumor immune responses
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Immune checkpoint combination: LGALS2 blockade may complement existing immune checkpoint inhibitors, potentially offering synergistic effects
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Biomarker development: Serum LGALS2 levels are significantly increased in metastatic breast cancer patients compared to healthy donors, suggesting potential as a diagnostic or prognostic biomarker
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Cross-cancer applications: Elevated LGALS2 expression appears to be a generalizable phenomenon across multiple cancer types, including testicular germ cell tumors, kidney renal clear cell carcinoma, and ovarian serous cystadenocarcinoma
What are common pitfalls in LGALS2 research and how can they be addressed?
| Challenge | Cause | Solution |
|---|---|---|
| Antibody specificity | Cross-reactivity with other galectins | Validate using LGALS2-KO controls; use multiple antibody clones |
| In vitro vs. in vivo discrepancy | Missing microenvironment interactions | Always confirm in vitro findings with in vivo models |
| Temporal dynamics | Time-dependent effects | Design longitudinal studies with multiple timepoints |
| Strain-dependent effects | Genetic background influences | Use multiple mouse strains; backcross to common background |
| Cell-specific expression | Heterogeneous expression patterns | Employ single-cell techniques and spatial transcriptomics |
How should researchers design controls for LGALS2 studies in mouse models?
Robust experimental design requires comprehensive controls:
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Genetic controls:
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Method-specific controls:
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Experimental validation:
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Confirm findings using multiple methodologies
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Include positive controls known to respond to manipulations
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Perform dose-response studies where applicable
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What technical innovations are advancing LGALS2 research in mouse models?
Recent technological advances have expanded research capabilities:
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Single-cell sequencing: Enabling cell-type-specific analysis of LGALS2 expression and effects, revealing previously undetected heterogeneity
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Spatial transcriptomics: Providing insights into LGALS2's role in tertiary lymphoid structures and the complexity of the tumor microenvironment at macro and micro levels
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CRISPR-based screens: In vivo screens under different immune pressures have characterized LGALS2 function among complex tumor microenvironments with various immune cell infiltrations
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Therapeutic antibodies: Development of specific inhibitory antibodies has enabled functional studies and therapeutic potential assessment
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Integrated computational approaches: Biological network-based computational strategies and machine learning methods have identified LGALS2 as a key marker within tertiary lymphoid structures
Product Science Overview
Galectin-2 is a member of the galectin family, which consists of carbohydrate-binding proteins known for their role in various cellular processes, including cell-cell adhesion, cell-matrix interactions, and intracellular signaling. Galectin-2, specifically, is a homodimeric protein with a molecular weight of approximately 14 kDa . It belongs to the S-type lectin family and contains a single carbohydrate-recognition domain (CRD) .
Galectin-2 is involved in modulating immune responses and has been shown to induce a proinflammatory phenotype in monocytes and macrophages . This protein plays a significant role in the regulation of inflammatory responses and has been associated with decreased collateral arteriogenesis in patients with obstructive coronary artery disease . The interaction of galectin-2 with CD14/TLR4 on monocytes leads to the induction of proinflammatory cytokines and inhibition of pro-arteriogenic factors .
Recombinant mouse galectin-2 is typically produced using human embryonic kidney (HEK293) cells . The gene encoding mouse galectin-2 is cloned into an expression vector, which is then transfected into HEK293 cells. The cells are cultured, and the recombinant protein is harvested from the culture medium. The protein is then purified using affinity chromatography techniques to ensure high purity and activity .
For large-scale production, recombinant mouse galectin-2 can be produced using bioreactor systems. The HEK293 cells expressing galectin-2 are grown in bioreactors under controlled conditions to maximize yield. The culture medium is continuously monitored and adjusted to maintain optimal growth conditions. After sufficient protein expression, the culture medium is collected, and the recombinant protein is purified using a series of chromatographic steps, including affinity chromatography and size-exclusion chromatography .
Galectin-2 binds to specific carbohydrate structures on the surface of cells, mediating various biological effects. The binding affinity of galectin-2 to its ligands is influenced by the presence of specific glycan structures. For instance, galectin-2 has been shown to interact with MUC5AC, a major gastric mucin glycoprotein, through affinity chromatography and subsequent LC-MS/MS analysis . This interaction highlights the specificity of galectin-2 for certain carbohydrate ligands and its potential role in modulating cellular functions through glycan recognition.
