Recombinant Human IL-17A/F Heterodimer

Recombinant Human IL-17A/F Heterodimer is a valuable tool for research applications due to its unique biological properties and relevance in immune regulation, inflammation, and disease pathogenesis. Here are key reasons to consider using it in your research:

1. Biologically Active and Intermediate Potency

• The IL-17A/F heterodimer is a biologically active cytokine that induces chemokine production and airway neutrophilia with intermediate potency between IL-17A (most potent) and IL-17F (least potent). This makes it ideal for studies where moderate inflammatory responses are desired, or for comparing the effects of different IL-17 family members.

2. Relevance in Inflammatory and Autoimmune Diseases

• IL-17A/F is upregulated in immune cells during inflammatory arthritis and contributes to disease severity. It is also implicated in autoimmune disorders and mucosal immunity, making it a relevant target for studying disease mechanisms and potential therapeutic interventions.

3. Structural and Functional Insights

• The heterodimer closely mimics both IL-17A and IL-17F, allowing for the study of receptor binding and signaling pathways. It can bind to both the "A-face" and "F-face" of the IL-17RA and IL-17RC receptors, enabling the formation of two topologically-distinct heterotrimeric complexes with potentially different signaling properties.

4. Induction of Pro-inflammatory Cytokines and Chemokines

• IL-17A/F stimulates the production of pro-inflammatory cytokines (IL-6, IL-1, TNF), granulopoietic factors (G-CSF, GM-CSF), antimicrobial factors (defensins, S100 proteins), chemokines (IL-8, CXCL1, CXCL5, CCL2, CCL7), and matrix metalloproteinases (MMP1, MMP3, MMP13). This broad spectrum of activity makes it useful for studying the inflammatory response and its regulation.

5. Applications in Cell Culture and Bioassays

• The recombinant protein is suitable for cell culture and bioassay applications, allowing researchers to investigate the effects of IL-17A/F on various cell types, including naïve CD4+ T cells, fibroblasts, and epithelial cells.

6. Research on Mucosal Immunity and Cancer

• Some studies suggest that IL-17A/F may be involved in mucosal immunity against bacterial infections and has a putative role in some autoimmune disorders and cancers. This makes it a valuable tool for research in these areas.

7. High Purity and Low Endotoxin Levels

• Recombinant Human IL-17A/F Heterodimer is typically available with high purity (≥95% by SDS-PAGE) and low endotoxin levels (≤1 EU/1 μg), ensuring reliable and reproducible results in your experiments.

8. Versatility in Experimental Design

• The heterodimer can be used in a variety of experimental setups, including ELISAs, Luminex assays, and other immunoassays, providing flexibility in your research approach.

In summary, Recombinant Human IL-17A/F Heterodimer is a versatile and biologically relevant cytokine that can provide valuable insights into the mechanisms of inflammation, immune regulation, and disease pathogenesis. Its intermediate potency and broad range of activities make it an excellent choice for a wide array of research applications.

Yes, recombinant Human IL-17A/F heterodimer can be used as a standard for quantification or calibration in ELISA assays, provided the ELISA is specifically designed to detect the IL-17A/F heterodimer and recognizes both natural and recombinant forms.

Key considerations and supporting details:

• ELISA specificity: The ELISA kit or assay must be validated for the detection of the IL-17A/F heterodimer, not just the IL-17A or IL-17F homodimers. Sandwich ELISA kits specifically designed for IL-17A/F heterodimer quantification use recombinant IL-17A/F as the standard and recognize both natural and recombinant forms.
• Standard preparation: Recombinant IL-17A/F heterodimer is typically reconstituted and serially diluted to generate a standard curve covering the assay’s dynamic range. For example, some kits use a range from 15.6 pg/mL to 500 pg/mL.
• Validation: The recombinant standard should be of high purity and bioactivity, with low endotoxin levels, and its concentration should be accurately determined. The standard curve must be freshly prepared for each assay to ensure reproducibility.
• Matrix compatibility: The standard can be used for quantification in various sample types, including cell culture supernatants, serum, and plasma, as long as the assay is validated for those matrices.
• Carrier protein: Some recombinant standards are supplied with carrier proteins (e.g., BSA) to enhance stability. If your assay is sensitive to carrier proteins, select a carrier-free formulation.

Important limitations:

• Do not use recombinant IL-17A/F heterodimer as a standard in ELISAs designed only for IL-17A or IL-17F homodimers, as cross-reactivity is minimal and quantification will be inaccurate.
• Always follow the manufacturer’s instructions for standard reconstitution, dilution, and storage to maintain assay accuracy and reproducibility.

In summary, recombinant Human IL-17A/F heterodimer is suitable as a standard for ELISA quantification when the assay is specifically validated for the heterodimer and recognizes both natural and recombinant forms.

Recombinant Human IL-17A/F Heterodimer has been validated for several key applications in published research, primarily in the study of immune and inflammatory responses.

Validated Applications:

• Cell-based Bioassays:
  Used to assess biological activity, such as induction of chemokine production, airway neutrophilia, and modulation of immune cell function. These assays have demonstrated that IL-17A/F induces chemokine production and airway neutrophilia with intermediate potency between IL-17A and IL-17F homodimers.

• Functional Assays:
  Employed to study the cytokine’s effects on various cell types, including fibroblasts, epithelial cells, endothelial cells, and immune cells. IL-17A/F has been shown to stimulate production of pro-inflammatory cytokines (e.g., IL-6, IL-8), neutrophil chemoattractants, and adhesion molecules, and to promote leukocyte recruitment.

• Cell Culture Studies:
  Utilized to investigate signaling pathways and cellular responses, such as activation of Erk1/2 and NF-κB pathways via the IL-17RA/RC receptor complex.

• Disease Models:
  Applied in preclinical models of immune-mediated inflammatory diseases (IMIDs), including rheumatoid arthritis (RA), inflammatory bowel disease (IBD), psoriasis, and asthma. IL-17A/F has been implicated in disease severity and progression, and its neutralization has been evaluated for therapeutic efficacy.

• Mucosal Immunity and Barrier Function:
  Studied for its role in protecting skin and mucosal barriers against infectious agents and in mucosal immunity against bacterial infections.

Representative Published Research:

• Induction of Chemokine Production and Airway Neutrophilia:
  IL-17A/F heterodimer induces chemokine production and airway neutrophilia in cell-based assays, with potency intermediate between IL-17A and IL-17F.

• Modulation of Epithelial and Smooth Muscle Cell Function:
  Shown to induce migration of human airway smooth muscle cells and regulate chloride-bicarbonate exchange in bronchial epithelial cells.

• Autoimmune and Inflammatory Disease Models:
  Used in studies of chronic mucocutaneous candidiasis, rheumatoid arthritis, and IBD to evaluate its contribution to disease severity and as a target for therapeutic intervention.

• Signaling Pathway Analysis:
  Validated for use in dissecting IL-17RA/RC-dependent signaling mechanisms in various cell types.

Summary Table of Validated Applications

Additional Notes:

• The heterodimer is often compared to IL-17A and IL-17F homodimers for potency and functional effects.
• It is typically used in in vitro and in vivo models to study cytokine signaling, immune cell recruitment, and inflammatory responses.

If you need protocols or more specific details on experimental design, please specify the application area.

To reconstitute and prepare Recombinant Human IL-17A/F Heterodimer protein for cell culture experiments, follow these steps:

• Reconstitution:
  Dissolve the lyophilized protein at a concentration of 100 μg/mL in sterile 4 mM HCl. For enhanced stability and to prevent adsorption, it is recommended to include at least 0.1% human or bovine serum albumin (HSA/BSA) in the reconstitution buffer if your application allows. If your protocol or supplier specifies a different buffer (e.g., sterile water), reconstitute at 0.1 mg/mL and further dilute as needed.

• Aliquoting:
  After reconstitution, gently mix the solution by pipetting (do not vortex). Divide into small aliquots to avoid repeated freeze-thaw cycles.

• Storage:
  Store aliquots at –20°C to –70°C for long-term use. For short-term use (up to 1 month), store at 2–8°C under sterile conditions. Avoid repeated freeze-thaw cycles to maintain protein integrity.

• Dilution for cell culture:
  Before adding to cell culture, dilute the stock solution in cell culture medium or a suitable buffer containing low endotoxin and a carrier protein (e.g., FBS or tissue culture grade BSA) to your desired working concentration. The optimal concentration should be determined empirically for each cell type and assay.

• General handling tips:

  • Briefly centrifuge the vial before opening to collect all material at the bottom.
  • Gently resuspend by pipetting along the vial wall.
  • Do not vortex, as this may denature the protein.

Summary Table: Preparation Protocol

Additional notes:

• If your protein is supplied carrier-free, adding BSA/HSA after reconstitution is recommended for long-term storage and stability.
• Always use sterile technique to avoid contamination.
• Check the specific supplier datasheet for any unique formulation or buffer requirements.

This protocol ensures optimal solubility, stability, and bioactivity of the IL-17A/F heterodimer for cell culture experiments.