COLEC11 Antibody
Description
Definition and Types of COLEC11 Antibodies
COLEC11 antibodies are produced in various formats, including monoclonal and polyclonal variants, tailored for specific experimental techniques. Key characteristics include:
| Source | Type | Reactivity | Applications |
|---|---|---|---|
| Abcam (ab278063) | Mouse monoclonal | Human, transfected cells | ELISA, WB, ICC/IF |
| Thermo Fisher (PA5-103231) | Rabbit polyclonal | Human, mouse, rat | WB, IHC, IF |
| Proteintech (15269-1-AP) | Rabbit polyclonal | Human, mouse, rat | WB, IHC, IF, IP, ELISA |
| Abbexa (P-42707) | Rabbit polyclonal | Human, mouse, rat | ELISA, WB, IHC-P |
These antibodies are affinity-purified and validated for specificity, with molecular weights ranging from 29–34 kDa, depending on the assay conditions .
Structure and Function of COLEC11 Protein
The COLEC11 protein (also known as collectin kidney 1, CL-K1) is a calcium-dependent lectin that binds high-mannose glycans and fucose residues on microbial surfaces or apoptotic cells . Its key functions include:
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Innate immunity: Activates the lectin complement pathway via MASP-1/3 recruitment, enhancing microbial clearance .
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Apoptosis regulation: Recognizes DNA on apoptotic cells, triggering complement activation .
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Embryogenesis: Guides neural crest cell migration during organogenesis, with mutations linked to 3MC syndrome .
Research Applications
COLEC11 antibodies are instrumental in studying:
3.1. Immunological Disorders
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Rheumatoid Arthritis (RA): COLEC11 deficiency exacerbates RA in mice by enhancing APC activation and Th1/Th17 responses, suggesting its immune-modulatory role .
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3MC Syndrome: Mutations in COLEC11 impair protein function, leading to facial dysmorphia and organ malformations .
3.2. Cancer Biology
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Tumor Microenvironment (TME): COLEC11 promotes tumor growth by reprogramming tumor-associated macrophages (TAMs) toward immunosuppressive M2 phenotypes. Depletion of COLEC11 reduces melanoma burden by enhancing cytotoxic T-cell infiltration .
3.3. Detection Methods
The antibody is commonly used in:
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ELISA: Quantifies COLEC11 levels in plasma (e.g., 2.1 μg/mL in healthy individuals) .
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Western Blot: Detects oligomeric forms (100–200 kDa) in CHO cell lysates .
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Immunohistochemistry: Visualizes COLEC11 in tissues like liver and adrenal glands .
Research Findings
Recent studies highlight COLEC11’s dual roles:
Product Specs
Target Background
- This study elucidates the role of COLEC11 and the molecular mechanisms involved in modulating retinal pigment epithelial cell phagocytosis and cytokine production. PMID: 28772263
- High COLEC11 expression has been associated with cardiovascular disease. PMID: 27341702
- The exclusion of the MASP1 and COLEC11 Loci in two individuals from different consanguineous families, along with the absence of mutations in four further individuals sequenced for both genes, suggests the possibility of further genetic heterogeneity in 3MC syndrome. PMID: 26789649
- Researchers have identified collectin CL-LK as a novel soluble C-type lectin capable of binding M. tuberculosis and characterized mycobacterial mannose-capped lipoarabinomannan as a primary ligand for CL-LK. PMID: 26173080
- Data indicate differences in the plasma concentrations of collectin liver 1 and collectin kidney 1, M-ficolin, and H-ficolin in systemic lupus erythematosus (SLE) patients compared to a group of healthy controls. PMID: 26154564
- The promoter polymorphism COLEC11-9570C>T (rs3820897) was associated with decreased levels of CL-K1. PMID: 25710878
- The sugar specificity of CL-K1 was established. PMID: 25912189
- These results suggest that specific diseases may affect CL-K1 synthesis in an organ-dependent manner, and elevated plasma CL-K1 levels are associated with the presence of DIC. PMID: 24474086
- Most plasma CL-K1 was found in complex with CL-L1 in a ratio suggesting a heteromeric subunit of 1 CL-L1 & 2 CL-K1 polypeptides. It associated with MASPs 1, 2 & 3. Upon binding mannan or DNA in the presence of MASP-2, the complex deposited C4b. PMID: 24174618
- Data suggest that collectin 11 (CL-11), potentially via complement, may play a role in response to particles and surfaces presenting extracellular DNA, such as apoptotic cells. PMID: 23954398
- Collectin-11 associates with all known MBL-associated serine proteases (MASP-1, MASP-2, and MASP-3), as well as the lectin complement pathway regulator MAP-1. PMID: 23220946
- Data indicate that the concentration of collectin kidney 1 (CL-K1, COLEC11) in plasma was 0.34 +/- 0.13 microg/ml, and that in mannan-binding lectin (MBL) was 1.72 +/- 1.51 microg/ml. PMID: 21893516
- These findings demonstrate a role for complement pathway factors in fundamental developmental processes and in the etiology of 3MC syndrome. PMID: 21258343
- COLEC11 plays a role in the activation of the complement system and in the defense against invading microorganisms. PMID: 20956340
Q&A
What is COLEC11 and why is it significant in cancer research?
COLEC11 (Collectin-11) is a member of the collectin family of C-type lectins that play multifunctional roles in homeostasis, host defense, and disease pathogenesis. Recent research has revealed its critical involvement in tumor biology, particularly in melanoma progression. COLEC11 has been found to promote cancer cell proliferation, angiogenesis, and the establishment of an immunosuppressive tumor microenvironment. Gene expression analysis demonstrates that COLEC11 expression is upregulated in human melanomas, with higher levels observed in metastatic melanomas compared to primary melanomas .
The significance of COLEC11 in cancer research is highlighted by studies using COLEC11-deficient (Colec11−/−) mice, which showed significantly reduced tumor burden, volume, and weight in both B16 and YUMM1.7 melanoma models compared to wild-type controls. This suggests that COLEC11 could be a potential therapeutic target in melanoma treatment .
What types of COLEC11 antibodies are available for research applications?
Several validated COLEC11 antibodies are available for research, with distinct characteristics suited for different experimental approaches:
The choice between these antibodies depends on the specific research application. The polyclonal antibody offers broader species reactivity and application range, while the monoclonal antibody provides higher specificity for human COLEC11 and is particularly suitable for immunocytochemistry applications .
How can COLEC11 antibodies be used to investigate tumor microenvironment changes?
COLEC11 antibodies can be effectively employed to analyze alterations in the tumor microenvironment (TME) using multiple methodologies:
Immunohistochemical Analysis
Use COLEC11 antibodies in combination with immune cell markers to characterize the TME. Research has shown that COLEC11 deficiency leads to significant changes in the cellular composition of tumor-infiltrating leukocytes, with higher proportions of lymphocytes (CD4+, CD8+, NK1.1+) and lower proportions of myeloid lineage cells (CD11b+, CD11b+Ly6G+, CD11b+Ly6G-Ly6G+) .
Co-staining Protocols
Implement double immunostaining using COLEC11 antibodies alongside:
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CD45 to identify immune-infiltrating versus non-immune cells
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CD3 and CD11b to assess T cell versus myeloid cell infiltration
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CD8 to evaluate cytotoxic T cell presence in the tumor core and edges
Research has demonstrated that COLEC11-deficient mice exhibit markedly increased CD8+ infiltrates in the tumor core and edges, suggesting enhanced anti-tumor immunity .
What are the recommended antibody dilutions and experimental conditions for different COLEC11 detection methods?
Optimal experimental conditions vary by application and specific antibody:
| Application | Recommended Dilution (Polyclonal) | Recommended Dilution (Monoclonal) | Critical Considerations |
|---|---|---|---|
| ELISA | 1 μg/ml | Not specified | Optimize coating conditions for recombinant COLEC11 |
| Western Blotting | 1/500 - 1/1000 | Not specified | Expected molecular weight: 29 kDa |
| IHC-P | 1/50 - 1/100 | Not specified | Heat-mediated antigen retrieval recommended |
| ICC/IF | Not specified | 1/500 | FITC-conjugate secondary antibody at 1/4000 dilution |
Researchers should note that optimal dilutions may require optimization for specific experimental systems. For immunocytochemistry applications, a protocol using the monoclonal antibody at 1/500 dilution with FITC-conjugated rabbit anti-mouse IgG secondary antibody (1/4000) has been successfully validated .
Genetic Controls
Compare staining patterns between wild-type tissues and Colec11−/− tissues. Research has shown that in melanomas from wild-type mice, CD45+ cells were positively stained with CL-11, whereas in Colec11−/− mice, CD45+ cells appeared negative for CL-11 staining .
RNA-Protein Correlation
Assess correlation between COLEC11 protein expression (detected by antibody) and mRNA expression (measured by qPCR). Studies have demonstrated that Colec11 mRNA was mainly detected in CD45+ cells, with very low levels in CD45- cells from wild-type mice, while no Colec11 mRNA was detected in CD45+ cells from Colec11−/− mice .
Multiple Antibody Validation
Cross-validate results using different antibodies targeting distinct epitopes of COLEC11 to confirm specificity.
How can COLEC11 antibodies be used to study its role in angiogenesis?
COLEC11 has been implicated in promoting angiogenesis within tumors. Researchers can investigate this process using:
Vessel Density Analysis
Combine COLEC11 antibodies with endothelial markers (CD31, von Willebrand factor) to quantify vascular development. Research has shown that CD31 and VWF were markedly reduced in melanomas of Colec11−/− mice compared with wild-type mice .
Differential Vessel Characterization
Use distinct endothelial markers to differentiate between vessel types:
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CD31 primarily detects larger vessels
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VWF predominantly stains smaller vessels and is released upon endothelial cell activation
This differential staining approach allows researchers to characterize the influence of COLEC11 on specific aspects of tumor vasculature development.
What approaches can be used to investigate COLEC11's role in macrophage polarization?
COLEC11 has been shown to influence tumor-associated macrophage (TAM) phenotype. Several methodologies can be employed:
Flow Cytometry-Based Cell Sorting
Isolate CD45+F4/80+ cells from tumors using FACS for downstream analysis of macrophage phenotype .
Transcriptomic Analysis
Perform RNA-Seq on sorted TAMs to identify differentially expressed genes. Studies have revealed 625 differentially expressed genes between TAMs from wild-type and Colec11−/− mice, primarily classified into "Immune system," "Signal transduction," "Signaling molecules and interaction," and "Cancer" pathways .
Marker Expression Analysis
Examine expression of:
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M1 markers: H2.Aa, H2.Ab1, H2.Eb1, H2.DMa, Fcgg4, CD64, Stat1, Stat2, Socs1, Ifng
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M2 markers: Cd36, Cd163, Cd206, Cd14, Il10
Research has shown that TAMs from Colec11−/− mice exhibit higher expression of M1 marker genes and lower expression of M2 marker genes compared to wild-type controls .
How can researchers assess COLEC11's impact on cytokine/chemokine profiles in the tumor microenvironment?
COLEC11 significantly influences the cytokine and chemokine profiles within tumors. Recommended approaches include:
qPCR Analysis
Measure intratumoral mRNA levels of relevant cytokines and chemokines. Studies have detected significantly higher levels of Ifng, Nos2, Il12, Ccl5, Cx3cl1, and Cxcl9 and lower levels of Arg1 in melanomas of Colec11−/− mice compared to wild-type mice .
Multiplex Cytokine Assays
Quantify protein levels of multiple cytokines simultaneously in tumor lysates or supernatants from tumor explant cultures.
Cell-Specific Cytokine Production
Combine flow cytometry with intracellular cytokine staining to identify specific cellular sources of cytokines within the tumor microenvironment.
What experimental designs are most effective for investigating COLEC11's direct effects on cancer cell proliferation?
COLEC11 has been demonstrated to directly stimulate melanoma cell proliferation. Effective experimental approaches include:
Proliferation Assays
Assess cancer cell proliferation in the presence of purified COLEC11 protein or conditioned media from COLEC11-expressing cells. Research has shown that CL-11 has a direct stimulatory effect on murine melanoma cell proliferation .
Ki67 Immunostaining
Quantify proliferating cells within tumors using Ki67 staining combined with CD45 to distinguish tumor cells from immune cells. Studies have demonstrated that Colec11−/− mice displayed a significant reduction of Ki67+ cells in the tumor core, with most Ki67+ staining not associated with CD45+ staining, confirming that Ki67+ cells are mainly tumor cells .
Signaling Pathway Analysis
Investigate COLEC11's effects on cancer-related signaling pathways. Research has shown that CL-11 can activate tyrosine kinase receptors (EGFR, HER3) and ERK, JNK, and AKT signaling pathways .
How can researchers explore the potential of COLEC11 as a therapeutic target in cancer?
Based on research showing that COLEC11 promotes tumor growth, several approaches can be used to investigate its therapeutic targeting:
Blocking Strategies
Utilize L-fucose to inhibit COLEC11 activity. Studies have demonstrated that blockade of CL-11 with L-fucose inhibited melanoma growth in mice .
Genetic Approaches
Use CRISPR/Cas9 or shRNA to knock down COLEC11 expression in cancer cells or tumor-associated immune cells.
Combination Therapy Models
Investigate the effects of COLEC11 inhibition in combination with immune checkpoint inhibitors or conventional cancer therapies.
What methodologies can be used to study COLEC11 expression in human cancer samples?
Several approaches can be employed to analyze COLEC11 expression in human cancer specimens:
Tissue Microarray Analysis
Perform immunohistochemistry using validated COLEC11 antibodies on tissue microarrays containing multiple cancer samples.
Gene Expression Analysis
Analyze COLEC11 gene expression in cancer datasets. Research has shown that COLEC11 gene expression is upregulated in human melanomas, with higher levels in metastatic melanomas compared to primary melanomas .
Single-Cell Analysis
Conduct single-cell RNA sequencing combined with protein detection to identify specific cell populations expressing COLEC11 within human tumors.
What are the storage and handling recommendations for maintaining COLEC11 antibody performance?
To ensure optimal antibody performance in research applications:
Storage Conditions
Store antibodies in aliquots at -20°C and avoid repeated freeze/thaw cycles. The polyclonal antibody from Abbexa is provided in PBS (pH 7.3) containing 0.05% Proclin-300 and 50% glycerol .
Working Dilution Preparation
When preparing working dilutions, use fresh, cold buffer and maintain samples on ice to preserve antibody activity.
Quality Control
Include appropriate positive and negative controls in each experiment to verify antibody performance. For COLEC11 antibodies, this might include wild-type versus Colec11−/− tissues or cells with known COLEC11 expression patterns.
