Recombinant Mouse RELMβ

Recombinant Mouse RELMβ is a valuable tool for research applications focused on mucosal immunology, metabolic regulation, and inflammatory disease models because it allows precise investigation of RELMβ’s biological functions in vitro and in vivo.

Key scientific reasons to use recombinant RELMβ include:

• Dissecting Gut Immune-Epithelial Circuits: RELMβ is a master regulator of type 2 immune programs in intestinal epithelial cells, influencing goblet cell and tuft cell gene expression, antimicrobial peptide production, and epithelial barrier integrity. Recombinant RELMβ enables controlled studies of these pathways, including its role in oral tolerance and food allergy models.

• Modeling Inflammatory Responses: RELMβ promotes macrophage foam cell formation and enhances inflammatory cytokine expression (TNFα, IL-1β, IL-6) via NF-κB signaling in macrophages. Recombinant protein can be used to stimulate immune cells in vitro or in vivo to study mechanisms of inflammation, atherosclerosis, and mucosal injury.

• Studying Host Defense and Tissue Repair: RELMβ recruits CD4+ T cells to sites of colonic infection, elevates IL-22 levels, and promotes intestinal epithelial cell proliferation, thereby limiting infection-associated tissue pathology. Recombinant RELMβ restores these protective effects in knockout models, making it essential for mechanistic studies of mucosal healing and immune cell trafficking.

• Metabolic Regulation: Administration of recombinant RELMβ impairs hepatic insulin sensitivity and glucose metabolism, providing a model for studying metabolic syndrome and diabetes-related pathways.

• Allergy and Airway Remodeling: RELMβ is upregulated in allergic airway inflammation and contributes to fibroblast proliferation, differentiation, and airway remodeling. Recombinant RELMβ can be used to probe these processes in pulmonary models.

Best practices for using recombinant RELMβ:

• Employ recombinant protein in dose-response and time-course experiments to define its direct effects on target cells or tissues.
• Use in knockout or transgenic mouse models to complement genetic studies and confirm specificity of observed phenotypes.
• Apply in organoid cultures or primary cell assays to dissect cell-type-specific responses.

Summary of applications:

• Elucidating RELMβ’s role in mucosal immunity, epithelial biology, and microbiome interactions.
• Investigating mechanisms of inflammation, tissue repair, and metabolic regulation.
• Validating therapeutic targets for allergy, colitis, atherosclerosis, and metabolic disease.

Using recombinant mouse RELMβ provides a controlled, reproducible means to study its multifaceted roles in health and disease, advancing both basic and translational research.

You can use recombinant Mouse RELMβ as a standard for quantification or calibration in your ELISA assays, provided that the recombinant protein is of high purity and its concentration is accurately determined. This is a common and accepted practice in ELISA assay development and quantification.

Key considerations and best practices:

• Purity and Quantification: The recombinant RELMβ should be highly purified, and its concentration should be measured precisely, ideally by methods such as HPLC or absorbance at 280 nm.
• Standard Curve Preparation: Prepare a standard curve using serial dilutions of the recombinant RELMβ in the same buffer or matrix as your samples to ensure accurate quantification.
• Parallelism and Dilution Linearity: Validate that the recombinant standard behaves similarly to endogenous RELMβ in your sample matrix. This is done by testing parallelism (comparing dilution curves of samples and standards) and dilution linearity (recovery of spiked recombinant protein in samples).
• Matrix Effects: If your samples are in complex matrices (e.g., serum, plasma), spike known amounts of recombinant RELMβ into these matrices to confirm that recovery is consistent and that there are no significant matrix effects.
• Antibody Recognition: Ensure that the antibodies used in your ELISA recognize both the recombinant and endogenous forms of RELMβ with similar affinity. Differences in post-translational modifications or folding between recombinant and native proteins can sometimes affect antibody binding.
• Controls: Include appropriate controls, such as endogenous positive controls, to confirm assay performance and specificity.

Summary Table: Recombinant Protein as ELISA Standard

Conclusion:
Using recombinant Mouse RELMβ as a standard is scientifically valid and widely practiced for ELISA quantification, as long as you validate the assay for parallelism, recovery, and antibody specificity. This ensures your quantification is accurate and reliable.

Recombinant Mouse RELMβ has been validated in published research for several key applications, primarily in studies of immune regulation, epithelial biology, inflammation, and host-microbe interactions.

Validated Applications in Published Research:

• In vivo functional studies:
  Recombinant RELMβ has been administered to mice to study its effects on gut immune-epithelial circuits, oral tolerance, and food allergy models. For example, in a mouse model of food allergy, recombinant RELMβ was injected to induce expression of antimicrobial and epithelial genes (e.g., Sprr2a, Reg3a, Reg3b) and to assess its role in immune regulation and anaphylaxis susceptibility.

• Ex vivo organoid stimulation:
  Recombinant RELMβ has been used to treat murine gut epithelial organoids derived from wild-type and genetically modified mice, as well as human intestinal organoids, to analyze direct effects on epithelial gene expression and immune signaling pathways.

• Enema delivery in knockout mice:
  In infectious colitis models, recombinant RELMβ was delivered via enema to Retnlb knockout mice to restore CD4+ T cell recruitment, increase IL-22 production, promote intestinal epithelial cell proliferation, and reduce mucosal pathology.

• Cell culture stimulation:
  Recombinant RELMβ has been applied to bronchial epithelial cells to study its ability to induce inflammatory cytokines (e.g., IL-8, IL-1β), particularly in the context of chronic obstructive pulmonary disease (COPD) and airway inflammation.

• Cancer cell assays:
  Overexpression and recombinant protein studies have been conducted to investigate the effects of RELMβ on invasion, metastasis, and angiogenesis in gastric cancer cell lines.

• Pulmonary inflammation models:
  Recombinant RELMβ has been detected and studied in bronchoalveolar lavage fluid following allergen challenge in mouse models of pulmonary inflammation, indicating its use in airway and lung research.

Summary Table:

Key experimental readouts include gene expression profiling (qPCR, RNA-seq), cytokine measurement (ELISA), immune cell recruitment (flow cytometry, histology), and functional assays for epithelial barrier and antimicrobial activity.

These applications demonstrate that recombinant mouse RELMβ is a versatile tool for dissecting its roles in mucosal immunity, inflammation, epithelial biology, and disease models.

To reconstitute and prepare Recombinant Mouse RELMβ protein for cell culture experiments, centrifuge the vial briefly to collect the lyophilized powder at the bottom, then add sterile distilled water or the recommended buffer to achieve a final concentration of 0.1–1.0 mg/mL, gently pipetting to dissolve without vortexing. For long-term stability, add a carrier protein such as 0.1% HSA or BSA, aliquot, and store at –20°C to –80°C, avoiding repeated freeze-thaw cycles.

Step-by-step protocol:

• Centrifuge the vial for 20–30 seconds before opening to ensure all powder is at the bottom.
• Add sterile distilled water or the buffer specified in the product datasheet. Most recombinant proteins are reconstituted in water unless otherwise indicated.
• Target concentration: Reconstitute to 0.1–1.0 mg/mL. For example, add 100–1000 µL water to 100 µg protein.
• Gently pipet the solution down the sides of the vial to dissolve the protein. Avoid vortexing or vigorous shaking to prevent denaturation.
• Allow to dissolve for 15–30 minutes at room temperature with gentle agitation.
• Add carrier protein (e.g., 0.1% HSA or BSA) if the protein will be stored long-term or at low concentrations to prevent adsorption and loss of activity.
• Aliquot the solution into working volumes (≥10 µL) to minimize freeze-thaw cycles.
• Storage: Store aliquots at –20°C to –80°C for long-term use. For short-term use (up to 1 week), store at 4°C.
• Avoid repeated freeze-thaw cycles to preserve protein integrity.

Additional notes for cell culture:

• Confirm the absence of endotoxin if using in sensitive cell assays.
• Dilute the reconstituted protein into cell culture medium immediately before use.
• If unsure about buffer compatibility, consult the product datasheet or perform a small-scale test.

Summary Table:

These steps ensure optimal recovery and stability of recombinant Mouse RELMβ for cell culture applications.