Recombinant Human Resistin

Recombinant Human Resistin is a valuable tool for research applications due to its well-documented roles in inflammation, immune modulation, metabolic regulation, and vascular biology. Here are several key reasons why you should consider using Recombinant Human Resistin in your research:

1. Study of Inflammatory Pathways

• Human resistin is a potent modulator of inflammation, promoting the production of pro-inflammatory cytokines such as TNF-α, IL-1β, IL-6, and chemokines like MIP-2.
• It enhances neutrophil activation, neutrophil extracellular trap (NET) formation, and recruitment, making it essential for studying acute inflammatory responses, including acute lung injury (ALI) and sepsis.
• Resistin can be used to investigate the mechanisms of TLR4 signaling and its impact on innate immunity.

2. Investigation of Innate Immunity and Host Defense

• Resistin exhibits broad-spectrum antimicrobial activity against both Gram-positive and Gram-negative bacteria, positioning it as a host defense peptide.
• It can be used to study the interplay between inflammation and antimicrobial responses, especially in models of infection or immune challenge.

3. Metabolic and Endothelial Dysfunction Studies

• Resistin is implicated in insulin resistance and glucose metabolism, making it relevant for research on obesity, type 2 diabetes, and metabolic syndrome.
• It promotes endothelial dysfunction by reducing eNOS expression, increasing oxidative stress, and upregulating adhesion molecules, which is useful for vascular biology and atherosclerosis research.

4. Cardiac and Pulmonary Disease Models

• Resistin is expressed in heart tissues and has been linked to right ventricular dysfunction and maladaptive remodeling in pulmonary hypertension.
• It can be used to model and study cardiac dysfunction, pulmonary vascular remodeling, and the effects of resistin-neutralizing therapies.

5. Standardization and Reproducibility

• Recombinant Human Resistin provides a consistent, purified protein source, ensuring reproducibility across experiments.
• It is available in various forms (e.g., carrier-free, with BSA) and can be used as an ELISA standard, in cell culture, or in animal models.

6. Therapeutic Target Validation

• Resistin is a potential therapeutic target in inflammatory, metabolic, and cardiovascular diseases.
• Using recombinant resistin allows for the evaluation of neutralizing antibodies, inhibitors, or other interventions aimed at modulating resistin activity.

7. Mechanistic Studies

• Resistin can be used to dissect signaling pathways, such as NF-κB, STAT3, and TBK1, and to study its interactions with receptors like TLR4.
• It enables the investigation of resistin’s dual roles in both promoting and limiting inflammation, depending on the context.

In summary, Recombinant Human Resistin is a versatile and biologically relevant protein for studying inflammation, immunity, metabolism, and vascular/cardiac dysfunction, making it an essential reagent for a wide range of research applications.

Yes, recombinant human resistin can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and properly validated for this purpose. Many commercial human resistin ELISA kits use recombinant resistin as their standard, and this is a common practice in quantitative immunoassays.

Key considerations and supporting details:

• Recombinant protein as standard: ELISA kits for human resistin routinely use recombinant resistin as the calibration standard to generate the standard curve for quantification. The recombinant protein is typically a dimeric protein of ~19.5 kDa, matching the native form found in human samples.

• Standard curve preparation: The standard curve in ELISA is constructed by serially diluting the recombinant resistin standard to known concentrations, which allows for the quantification of resistin in unknown samples by interpolation.

• Purity and validation: It is essential that the recombinant resistin used as a standard is of high purity and has been validated for use in ELISA. The protein should be correctly folded and biologically active, as improper folding or degradation can affect antibody recognition and quantification accuracy.

• Matrix effects: When using recombinant resistin as a standard, it is important to prepare the standard dilutions in the same buffer or matrix as your samples (e.g., serum, plasma, or assay diluent) to minimize matrix effects and ensure accurate quantification.

• Documentation: Always refer to the specific ELISA kit protocol or manufacturer’s recommendations regarding the use of recombinant standards, as some kits may require specific formulations or additives (such as BSA) for optimal performance.

Summary Table: Use of Recombinant Human Resistin as ELISA Standard

In summary: As long as your recombinant human resistin is of high quality and appropriately validated, it is suitable for use as a standard in ELISA assays for quantification or calibration purposes. Always ensure compatibility with your specific assay system and follow best practices for standard preparation.

Recombinant Human Resistin has been validated for a variety of applications in published research, primarily focusing on its functional and biochemical roles. The key applications include:

• Functional Assays: Resistin has been used in studies to assess its effects on cell proliferation, migration, and capillary-like tube formation in endothelial cells, as well as its impact on inflammation, immune cell activation, and antimicrobial activity.
• Western Blot: Used to detect and quantify resistin protein expression in various tissues and cell types.
• ELISA (Enzyme-Linked Immunosorbent Assay): Employed for measuring resistin levels in biological samples, often as a standard or in immunoassays.
• Cell and Tissue Culture: Applied in studies investigating resistin's effects on different cell types, including endothelial cells, chondrocytes, and immune cells.
• Immunoassays: Utilized in immuno-staining and antibody-based detection methods to study resistin expression and localization in tissues.
• Biochemical and Biophysical Studies: Used in experiments to analyze resistin's structure, including its tendency to form disulfide-linked dimers and its conformational changes.

These applications highlight the versatility of recombinant human resistin in both basic research and clinical studies, particularly in the fields of cardiovascular disease, inflammation, and innate immunity.

To reconstitute and prepare Recombinant Human Resistin protein for cell culture experiments, follow these best-practice steps:

1. Centrifuge the vial: Before opening, briefly centrifuge the lyophilized protein vial (3000–3500 rpm for 5 minutes or a quick microcentrifuge spin) to ensure all powder is at the bottom and not lost when opening.

2. Reconstitution:

   • Use sterile, high-purity water (e.g., 18 MΩ-cm H₂O or sterile deionized water) as the initial solvent.
   • Typical reconstitution concentration is 0.1–1.0 mg/mL (100–1000 µg/mL), with 100 µg/mL being commonly recommended.
   • Gently add the water to the vial. Do not vortex; instead, gently swirl or pipette up and down to dissolve. Let the vial sit at room temperature for 15–30 minutes to ensure complete dissolution.
   • If the protein is difficult to dissolve, incubate at 4°C overnight.

3. Dilution for cell culture:

   • After reconstitution, dilute the protein to the desired working concentration using your cell culture medium or PBS.
   • Add a carrier protein (such as 0.1% BSA, 10% FBS, or 5% HSA) to the dilution buffer to prevent adsorption and loss of protein activity, unless your experiment requires serum-free or animal protein-free conditions.
   • For serum-free or animal-free applications, use trehalose as a stabilizer.

4. Aliquot and storage:

   • Prepare aliquots sized for single use to avoid repeated freeze-thaw cycles, which can denature the protein and reduce activity.
   • Store aliquots at 4°C for up to one week for short-term use, or at -20°C to -80°C for long-term storage (up to 3–6 months), ideally in the presence of a carrier protein or 5–50% glycerol.
   • Avoid storing at concentrations above 1 mg/mL to prevent solubility issues.

5. Handling precautions:

   • Do not vortex or shake vigorously at any step to avoid denaturation.
   • Confirm protein integrity and concentration if needed by SDS-PAGE or other analytical methods.

Summary protocol:

• Centrifuge vial → Add sterile water (0.1–1.0 mg/mL) → Gently dissolve → Dilute with medium + carrier protein → Aliquot → Store at 4°C (short-term) or -20°C/-80°C (long-term).

Note: Always consult the specific product’s Certificate of Analysis or datasheet for any manufacturer-specific recommendations, as buffer composition and pH requirements may vary.

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