Recombinant Human IL-17B

Recombinant Human IL-17B is used in research applications to investigate its roles in inflammation, cancer biology, immune signaling, and therapeutic resistance. Its utility stems from its ability to model and dissect the biological effects of the native cytokine in controlled experimental systems.

Key scientific applications and rationales include:

• Studying Inflammatory Pathways: IL-17B binds to the IL-17RB receptor, inducing the production of inflammatory cytokines and promoting infiltration of immune cells, making it a valuable tool for dissecting inflammatory signaling cascades and their downstream effects.

• Cancer Research: Recombinant IL-17B is widely used to model its role in tumorigenesis, cancer stem cell self-renewal, and metastasis. It has been shown to:

  • Promote tumor cell growth, migration, and invasion in vitro and in vivo.
  • Enhance cancer stem cell properties, such as sphere formation and expression of stemness markers (e.g., Lgr5, CD133).
  • Induce resistance to chemotherapeutic agents (e.g., paclitaxel) via activation of the ERK1/2 pathway and upregulation of anti-apoptotic proteins, providing a model to study mechanisms of drug resistance and potential therapeutic interventions.

• Autoimmunity and Tissue Repair: IL-17B is implicated in autoimmune and inflammatory diseases, such as arthritis, where it is upregulated in diseased tissue and exacerbates pathology. It is also involved in tissue repair processes, particularly in nervous tissue and cartilage.

• Molecular and Cellular Mechanisms: Recombinant IL-17B enables the study of:

  • Signal transduction via the IL-17RB receptor, including downstream pathways like NF-κB and MAPK.
  • Regulation of mRNA stability and translation of inflammatory genes.
  • Crosstalk with other cytokines and immune cell types, such as neutrophils and macrophages.

• Therapeutic Target Validation: By using recombinant IL-17B in cell-based assays and animal models, researchers can validate the IL-17B/IL-17RB axis as a potential therapeutic target for cancer, inflammatory diseases, and drug resistance.

Summary of best practices:

• Use recombinant IL-17B in bioassays to stimulate target cells and measure cytokine production, proliferation, migration, or resistance phenotypes.
• Employ it in animal models to assess its effects on tumor growth, metastasis, and response to therapy.
• Combine with pathway inhibitors or receptor knockdown to dissect specific signaling mechanisms.

In conclusion, recombinant human IL-17B is a versatile tool for elucidating the molecular and cellular functions of IL-17B in health and disease, particularly in the contexts of inflammation, cancer progression, and therapeutic resistance.

Yes, recombinant human IL-17B can be used as a standard for quantification and calibration in ELISA assays. This is a well-established application for this recombinant protein in immunoassay development.

Suitability as an ELISA Standard

Recombinant human IL-17B is specifically designed and recommended for use as an ELISA standard. The protein is produced with high biological activity and consistency, making it suitable for generating standard curves and calibrating sandwich ELISA assays for measuring both natural and recombinant IL-17B in biological samples.

Protein Characteristics

The recombinant IL-17B protein is approximately 20 kDa and is typically produced as a glycosylated cytokine. When selecting a formulation, recombinant IL-17B supplied with bovine serum albumin (BSA) is specifically recommended for use in ELISA applications as a standard. This formulation helps maintain protein stability and prevents non-specific binding during the assay.

Application in Quantification

Recombinant IL-17B standards are used to establish dose-response curves across defined concentration ranges. For example, typical detection ranges for IL-17B ELISA assays span from approximately 15.63 pg/mL to 1000 pg/mL or higher, depending on the specific assay configuration. The recombinant protein allows you to generate linear standard curves that enable accurate interpolation of sample concentrations.

Practical Considerations

When using recombinant IL-17B as a standard, ensure that your capture and detection antibodies are validated to recognize the recombinant form. The protein should be stored according to manufacturer specifications and reconstituted properly before use to maintain activity and reproducibility across multiple assay runs.

Recombinant Human IL-17B has been validated for several key applications in published research, primarily focusing on its role in cancer biology, inflammation, and stem cell regulation. The main applications supported by peer-reviewed studies include:

1. Cancer Stem Cell (CSC) Self-Renewal and Tumorigenesis

   • Recombinant IL-17B (rIL-17B) has been shown to promote the sphere-formation ability of cancer stem cells in vitro, indicating its role in enhancing stemness and self-renewal capacity.
   • In vivo studies demonstrate that rIL-17B enhances tumor growth and metastasis, particularly in gastric cancer (GC) models.
   • Silencing IL-17RB (the receptor for IL-17B) abrogates these effects, confirming the specificity of the IL-17B/IL-17RB signaling pathway in cancer progression.
   • Reference: Cell Reports, 2022; Oncogene, 2021; PMC7994204

2. Regulation of Autophagy and Ubiquitination

   • IL-17B signaling through IL-17RB regulates Beclin-1 ubiquitination and autophagy activation in cancer cells, suggesting a role in cellular homeostasis and survival mechanisms.
   • Reference: Cell Reports, 2022; Oncogene, 2021

3. Inflammatory Cytokine Production and Immune Cell Infiltration

   • IL-17B binds to IL-17RB and induces the production of inflammatory cytokines and the infiltration of inflammatory immune cells, highlighting its involvement in inflammatory responses.
   • Reference: R&D Systems; Clin. Exp. Immunol., 2011

4. Bone Growth and Development

   • IL-17B is expressed in bone collars and chondrocytes during development and after bone fracture, indicating a role in skeletal system regulation and bone growth.
   • Reference: R&D Systems

5. Cancer Cell Survival, Proliferation, and Migration

   • In mouse models, IL-17B signaling through IL-17RB directly promotes cancer cell survival, proliferation, and migration, and induces resistance to therapy.
   • Reference: The Emerging Role of the IL-17B/IL-17RB Pathway in Cancer, 2020

6. Bioassays and Functional Studies

   • Recombinant IL-17B is widely used in bioassays to study its effects on cell signaling, cytokine production, and cell migration.
   • Reference: R&D Systems; Abcam; Miltenyi Biotec

These applications underscore the importance of recombinant human IL-17B in both basic research and translational studies, particularly in the fields of oncology, immunology, and regenerative medicine.

To reconstitute and prepare Recombinant Human IL-17B protein for cell culture experiments, dissolve the lyophilized protein in an appropriate sterile buffer, typically at a concentration of 100–200 μg/mL, and include a carrier protein such as BSA if required for stability.

Detailed protocol:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Reconstitution buffer:
  • For most applications, use sterile PBS (phosphate-buffered saline).
  • If the protein is carrier-free, use sterile PBS alone.
  • If stability is a concern or for dilute solutions, add 0.1% BSA (bovine serum albumin) or human serum albumin to the PBS.
  • Alternatively, some protocols recommend sterile water for initial reconstitution, especially for His-tagged proteins, at concentrations not less than 200 μg/mL.
• Reconstitution concentration:
  • Commonly, reconstitute to 100 μg/mL in PBS (with or without 0.1% BSA).
  • For certain preparations, reconstitute to 200 μg/mL in sterile water.
  • For ELISA or standard curve preparation, concentrations may vary (e.g., 10,000 pg/mL for standards).
• Dissolution:
  • Gently pipette up and down or swirl to dissolve.
  • Incubate at room temperature for at least 20 minutes to ensure complete dissolution.
• Aliquoting and storage:
  • Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles.
  • Store aliquots at 2–8°C for short-term use (2–7 days).
  • For long-term storage, freeze at –20°C or below.
  • If using carrier-free protein, avoid storing at very dilute concentrations to prevent adsorption to tube walls.
• Preparation for cell culture:
  • Dilute the stock solution into cell culture medium immediately before use to the desired working concentration (often in the range of ng/mL to μg/mL, depending on your assay).
  • If using serum-free media, ensure the presence of a carrier protein to stabilize the cytokine.
  • Filter sterilize if necessary, using a 0.2 μm filter, especially if reconstituted in water or if sterility is uncertain.

Key points:

• Use sterile technique throughout.
• Avoid repeated freeze-thaw cycles.
• Always check the specific product datasheet for any unique requirements, as expression system or formulation may affect optimal reconstitution conditions.

Summary Table:

These guidelines ensure optimal solubility, stability, and bioactivity of recombinant IL-17B for cell culture applications.