Il1rl2 Antibody

What is IL1RL2 and why is it an important research target?

IL1RL2, also known as IL-36R or IL-1Rrp2, is a member of the interleukin-1 receptor family. This protein has gained significant research attention because:

• It forms part of a cytokine receptor gene cluster on chromosome 2q12, alongside IL1R1, IL1R2, IL1RL1, and IL18R1

• It functions primarily as a mediator of inflammatory responses

• It has demonstrated elevated expression in multiple cancer types, including bladder, colorectal, breast, gastric, and lung cancers

• It may play a regulatory role in inflammatory bowel diseases, suggesting its importance in intestinal homeostasis

Research on IL1RL2 is particularly valuable because it provides insights into the connection between chronic inflammation and cancer development, with inflammation being recognized as a hallmark of cancer progression .

How can I distinguish between different isoforms of IL1RL2 when conducting antibody-based detection?

At least three isoforms of IL1RL2 are known to exist, which presents both challenges and opportunities for specific detection:

• Most commercially available antibodies detect all three isoforms, such as those developed by Thermo Fisher Scientific

• For isoform-specific detection, examine the epitope region targeted by the antibody - those developed against specific amino acid sequences may offer greater specificity

• Western blotting with controls is recommended to verify which isoforms are detected in your experimental system

• If different isoforms must be distinguished, consider using antibodies targeting unique regions of each isoform, or employ RT-qPCR with isoform-specific primers as a complementary approach

When validating antibody specificity, research has shown that IL1RL2 antibodies typically do not cross-react with IL-1R or IL-1RL1, providing confidence in target-specific detection .

What are the recommended positive controls for validating IL1RL2 antibody performance?

For proper validation of IL1RL2 antibodies, the following positive controls are recommended based on established research protocols:

• Human small intestine tissue lysate has been identified as an effective positive control

• BLCA cell lines (bladder cancer) demonstrate higher IL1RL2 mRNA expression compared to SV-HUC-1 cells (normal urothelial cells)

• Validation experiments should include both western blotting and RT-qPCR to confirm antibody specificity at both protein and transcript levels

• For immunohistochemistry applications, BLCA tissues have shown high IL1RL2 expression compared to adjacent normal tissues, making them suitable positive controls

Research has shown significant upregulation of IL1RL2 protein expression in both cancer cell lines and tissues, providing useful benchmarks for antibody validation .

What are the optimal dilutions and conditions for using IL1RL2 antibodies in different applications?

Optimal conditions vary based on application and specific antibody properties. Based on published protocols and manufacturer recommendations:

It's important to note that optimal dilutions should ultimately be determined by each researcher through titration experiments in their specific experimental system . Buffer composition (PBS, pH 7.3, containing 0.02% sodium azide, 50% glycerol) has been shown to maintain antibody stability and performance .

How should I design RT-qPCR experiments to complement IL1RL2 antibody studies?

RT-qPCR serves as a valuable complement to antibody-based detection of IL1RL2. Design considerations include:

• Multiple studies have successfully used RT-qPCR to assess IL1RL2 mRNA levels in cell lines and tissues, particularly when examining differential expression between normal and cancer cells

• Primer design should target conserved regions across all isoforms for total IL1RL2 expression, or unique regions for isoform-specific detection

• The TCGA (The Cancer Genome Atlas) database can be used to validate expression patterns observed in experimental systems

• Appropriate reference genes must be selected based on the experimental context to ensure accurate normalization

In bladder cancer research, RT-qPCR revealed significantly higher IL1RL2 mRNA expression in tumor cell lines compared to SV-HUC-1 cells, which was subsequently confirmed by western blotting for protein levels .

What experimental approaches can be used to study IL1RL2 function beyond antibody-based detection?

Beyond antibody-based detection, several complementary approaches have proven effective:

• CRISPR/Cas9-mediated genetic engineering to examine loss-of-function phenotypes, as used in studies investigating IL1RL2 deficiency in macrophages and epithelial cells

• Induced pluripotent stem cell (iPSC) differentiation toward relevant cell types (macrophages, colonic organoids) to study developmental impacts

• Co-expression network analysis and functional enrichment to understand biological pathways and functions associated with IL1RL2

• Protein-protein interaction networks constructed using interaction gene search tools to identify molecular partners

• Analysis of signaling pathway activation (NFκB and MAPK) in response to IL1RL2 modulation

These approaches provide a comprehensive understanding of IL1RL2 function in different cellular contexts and disease states.

How can I address inconsistent antibody performance in IL1RL2 detection across different sample types?

Inconsistent antibody performance is a common challenge with IL1RL2 detection. Methodological solutions include:

• Verify epitope conservation across species if working with non-human samples, as antibody reactivity varies (e.g., some antibodies react with human and mouse, while others are human-specific)

• Optimize protein extraction protocols based on sample type - detergent composition significantly affects membrane protein solubilization

• For fixed tissues or cells, test multiple antigen retrieval methods as epitope accessibility may be differentially affected

• When switching between applications (e.g., WB to IHC), antibody performance may vary significantly due to differences in protein conformation

• Consider post-translational modifications that may mask epitopes in certain tissues or disease states

The observed molecular weight of IL1RL2 (65 kDa) should be consistent across properly prepared samples, serving as a quality control benchmark .

How should I interpret IL1RL2 expression data in relation to disease progression in cancer studies?

Research on IL1RL2 in cancer contexts suggests:

• In bladder cancer, IL1RL2 is highly expressed in tumor tissues compared to adjacent normal tissues, as demonstrated through IHC analysis of 17 pairs of samples

• This elevated expression correlates with clinical and pathological features, which can be analyzed using tables like those in the bladder cancer study showing relationships with patient variables (age, sex, BMI, smoking status, etc.)

• When interpreting expression data, consideration should be given to the three known isoforms and their potentially distinct functions

• Co-expression network analysis can help identify genes with expression patterns correlated with IL1RL2, providing insights into affected pathways

The table below from bladder cancer research illustrates how IL1RL2 expression correlates with patient variables:

Such data should be interpreted in the context of sample size, statistical power, and potential confounding factors .

What factors affect antibody specificity in IL1RL2 detection, and how can they be mitigated?

Several factors influence IL1RL2 antibody specificity:

• Epitope selection is critical - antibodies targeting different amino acid sequences of IL1RL2 (e.g., AA 1-116, AA 20-118, AA 107-156, AA 257-286) demonstrate variable specificity profiles

• Purification methods affect specificity - antibodies purified through protein A columns followed by peptide affinity purification show enhanced specificity

• Host species impacts cross-reactivity - rabbit-derived polyclonal antibodies often show broader species reactivity than mouse monoclonals

• Validation across multiple techniques (WB, ELISA, IHC) confirms consistent target recognition

To mitigate specificity issues:

• Use antibodies with validated specificity through knockout or knockdown controls

• Implement competitive binding assays with the immunizing peptide

• Compare results across multiple antibodies targeting different epitopes of IL1RL2

• Verify findings with complementary techniques like RT-qPCR

How can IL1RL2 antibodies be utilized in therapeutic development research?

IL1RL2 antibodies have potential applications in therapeutic research:

• In combination therapy models, IL1RL2 targeting could be explored alongside IL-2 administration, similar to approaches in HIV-1 therapy where IL-2 was used in combination with therapeutic immunization

• For immunotherapy development, IL1RL2 antibodies can help monitor receptor expression changes in response to treatment

• Humanized antibodies against IL1RL2 could be developed for direct therapeutic application, especially in cancers where it shows elevated expression

• In inflammatory bowel disease research, where IL1RL2 mutations have been identified as risk factors for very early onset IBD (VEO-IBD), antibodies can help characterize patient-specific expression patterns

Research has shown reduced activation of NFκB and MAPK signaling pathways in patient-derived macrophages with IL1RL2 mutations, suggesting potential therapeutic targets in this pathway .

What approaches enable the design of antibodies with customized specificity profiles for IL1RL2 research?

Advanced approaches to antibody design include:

• Biophysics-informed models trained on experimentally selected antibodies can identify distinct binding modes associated with specific ligands

• Phage display experiments involving antibody selection against combinations of closely related ligands can generate highly specific variants

• Computational prediction and generation of antibody variants not present in initial libraries can create antibodies with tailored specificity profiles

• Experimental validation of computationally designed antibodies confirms their ability to discriminate between very similar epitopes

This advanced approach has applications not only in designing antibodies with specific properties but also in mitigating experimental artifacts and biases in selection experiments .

How can IL1RL2 antibodies be effectively employed in emerging single-cell analysis techniques?

Integration of IL1RL2 antibodies into single-cell technologies:

• For single-cell protein profiling, antibodies conjugated with heavy metals or fluorophores can be used in mass cytometry or flow cytometry to examine IL1RL2 expression heterogeneity within tissues

• In spatial transcriptomics combined with protein detection, IL1RL2 antibodies can reveal the spatial distribution of the receptor in the tissue microenvironment

• For multi-omics approaches, antibody-based sorting of IL1RL2-positive cells followed by RNA-seq or ATAC-seq can reveal transcriptional and epigenetic features associated with IL1RL2 expression

• In tumor microenvironment studies, IL1RL2 antibodies can help identify specific cell populations and their interactions in the inflammatory milieu

These applications require highly specific antibodies validated for the particular single-cell technology being employed, with careful attention to background signal and non-specific binding.

What are the emerging research areas for IL1RL2 antibodies beyond current applications?

Several promising research frontiers await further exploration:

• Investigation of IL1RL2's role in the tumor microenvironment and its potential as an immune checkpoint target, given its involvement in inflammatory responses

• Development of antibody-drug conjugates targeting IL1RL2 for directed therapy in cancers with high expression

• Exploration of IL1RL2's function in intestinal homeostasis, particularly in the context of inflammatory bowel diseases where compound heterozygous mutations have been identified

• Use of advanced biophysics-informed models to design antibodies with unprecedented specificity for IL1RL2, enabling more precise targeting

• Integration of IL1RL2 antibodies in multi-parameter imaging techniques to understand its spatial distribution in healthy and diseased tissues

The connection between IL1RL2 and both inflammation and cancer suggests untapped potential for therapeutic intervention in multiple disease contexts .

How might IL1RL2 antibody research contribute to understanding the link between inflammation and cancer?

Given that chronic inflammation is recognized as a hallmark of cancer, IL1RL2 research bridges these fields:

• IL1RL2's role as a mediator of inflammatory responses and its high expression in multiple cancer types suggests mechanistic connections worth exploring

• Antibody-based studies can help delineate how IL1RL2 signaling influences the tumor microenvironment and immune cell infiltration

• Investigation of IL1RL2's interaction with its agonists (IL-36α, IL-36β, and IL-36γ) in the cancer context could reveal new therapeutic targets

• Research on IL1RL2's pro-metastatic effects, as observed in colorectal cancer, could extend to other cancer types

• The association between IL1RL2 expression and patient prognosis in multiple cancers suggests its value as a biomarker for inflammation-driven cancer progression