Recombinant Human Chemerin

Recombinant Human Chemerin is used in research applications to study its roles in inflammation, metabolism, immune cell recruitment, and cancer biology, as well as to model its therapeutic potential in disease contexts. Its recombinant form provides a standardized, bioactive protein for controlled experimental manipulation.

Key reasons to use recombinant human chemerin in research include:

• Immune Cell Recruitment and Inflammation: Chemerin is a chemoattractant that recruits immune cells such as dendritic cells and macrophages to sites of inflammation or tissue injury, making it valuable for studying immune responses and inflammatory diseases.
• Metabolic Regulation: As an adipokine, chemerin regulates adipogenesis and glucose metabolism, making it relevant for research into obesity, diabetes, and metabolic syndrome.
• Cancer Research: Chemerin has demonstrated both pro- and anti-tumor effects, depending on the context. It can inhibit tumor growth and metastasis in some cancers by modulating immune cell infiltration and angiogenesis, or promote invasion in others via specific signaling pathways. Recombinant chemerin allows for direct testing of these effects in vitro and in vivo.
• Neuroprotection: Recombinant human chemerin has shown neuroprotective effects in rodent models of stroke, reducing brain damage and improving neurological outcomes, suggesting potential for translational research in neuroprotection and stroke therapy.
• Standardization for Assays: Recombinant chemerin is used as a standard in ELISA, Western blot, and bioassays, ensuring reproducibility and quantification in experimental protocols.
• Mechanistic Studies: It enables precise dissection of chemerin’s signaling pathways (e.g., via CMKLR1, RhoA/ROCK1, PTEN-AKT) in cellular models, supporting mechanistic studies of cell migration, invasion, and cytokine production.

Applications include:

• Cell culture experiments to assess chemerin’s effects on cell migration, proliferation, and signaling.
• Animal models to study chemerin’s role in disease progression or therapy.
• Use as a standard or control in immunoassays (ELISA, Western blot).
• Investigation of chemerin’s impact on metabolic, inflammatory, or oncogenic pathways.

Summary:
Using recombinant human chemerin provides a reliable, bioactive tool to elucidate its diverse biological functions and therapeutic potential in inflammation, metabolism, cancer, and neuroprotection, while enabling standardized and reproducible research protocols.

Yes, recombinant human Chemerin can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately determined. This is a common and accepted practice in both commercial ELISA kits and custom assay development.

Key points and best practices:

• Recombinant Chemerin is routinely used as a standard in commercial ELISA kits for human Chemerin quantification. Multiple kit instructions and technical datasheets specify that their calibration curves are generated using recombinant human Chemerin protein.
• Purity and quantification: The recombinant protein used as a standard should be highly purified and its concentration accurately measured, ideally by methods such as HPLC or absorbance at 280 nm. Impurities or inaccurate quantification can lead to errors in your standard curve and sample measurements.
• Standard curve preparation: Prepare a serial dilution of the recombinant Chemerin in the same buffer or matrix as your samples to minimize matrix effects. The standard curve should cover the expected range of Chemerin concentrations in your samples.
• Validation: If you are developing your own ELISA or using a recombinant standard not provided with a kit, validate the standard curve for linearity, accuracy, and reproducibility. Compare recovery and parallelism with native Chemerin in biological samples if possible.
• Documentation: Many ELISA kit manuals explicitly state that their standards are recombinant human Chemerin, and some even specify the expression system (e.g., E. coli-derived).

Summary Table: Use of Recombinant Chemerin as ELISA Standard

References to protocols and best practices:

• "A purified protein should be used to prepare the standard curve. Otherwise, use a recombinant protein which can be semi-purified in the lab and measure the concentration with HPLC".
• "The standards are prepared from recombinant human Chemerin".
• "Applications: ELISA (Standard)" for recombinant human Chemerin protein.
• "This DuoSet is calibrated against a highly purified E. Coli-expressed recombinant Human Chemerin".
• "Lyophilized recombinant human Chemerin standard".

In summary: As long as your recombinant human Chemerin is of high purity and accurately quantified, it is appropriate and standard practice to use it as a calibrator or standard for quantification in ELISA assays.

Recombinant Human Chemerin has been validated for several applications in published research, primarily in bioassays, ELISA (as a standard), and Western blot. It has also been used in functional studies involving cell signaling, migration, and disease models.

Key validated applications include:

• Bioassays:
  Recombinant human chemerin has been widely used in cell-based bioassays to study its effects on various cell types, including:

  • Inhibition of neoangiogenesis and antitumoral effects in cancer models.
  • Promotion of inflammation in glomerular endothelial cells relevant to diabetic nephropathy.
  • Investigation of cell migration, invasion, and EMT (epithelial-mesenchymal transition) in cancer cells.
  • Modulation of insulin signaling, glucose uptake, and stress kinase activation in human skeletal muscle cells.
  • Neuroprotection in rodent models of stroke, both as pre- and post-ischemic intervention.

• ELISA (Standard):
  Recombinant chemerin is validated as a standard for quantifying chemerin levels in ELISA assays, enabling measurement of endogenous chemerin in biological samples.

• Western Blot:
  Used as a positive control or standard for detecting chemerin protein in tissue or cell lysates by immunoblotting.

• Other Functional Assays:

  • Chemoattraction assays, such as migration of human immature dendritic cells.
  • Surface-enhanced Raman scattering for disease biomarker detection.
  • Studies on metabolic syndrome, diabetes, atherosclerosis, and cardiovascular diseases, often involving cell culture or animal models to assess chemerin’s physiological and pathological roles.

Summary Table of Validated Applications

Additional Notes:

• Most studies use recombinant human chemerin in in vitro cell culture systems or in vivo animal models.
• The protein is often used to probe signaling pathways, cellular responses, and disease mechanisms.
• It is important to select the appropriate format (carrier-free or with BSA) depending on the application, especially for cell culture or as an ELISA standard.

If you need protocol-specific details or information on a particular disease model or assay, please specify the context.

To reconstitute and prepare Recombinant Human Chemerin protein for cell culture experiments, dissolve the lyophilized protein in sterile distilled water to a final concentration of 0.1 mg/mL. The protein is typically supplied lyophilized from a buffer containing 0.1–0.2% trifluoroacetic acid (TFA).

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all material is at the bottom.
• Add sterile distilled water to achieve the desired concentration (commonly 0.1 mg/mL; for example, add 100 µL water to 100 µg protein).
• Gently pipette the solution down the sides of the vial to dissolve the protein. Avoid vigorous mixing or vortexing to prevent foaming and protein denaturation.
• Allow the protein to dissolve for 15–30 minutes at room temperature with gentle agitation.
• If necessary, wash the vial with additional water to recover any remaining protein.
• Once fully dissolved, filter-sterilize the solution using a 0.2 µm filter if required for cell culture applications.
• Aliquot the solution to avoid repeated freeze-thaw cycles and store at −20°C for long-term use or at 4°C for short-term use (up to one week).

Additional considerations:

• If the protein is supplied in TFA, you may need to buffer exchange or dilute the solution in cell culture medium to minimize TFA concentration before adding to cells.
• Always consult the specific product datasheet for recommended reconstitution buffer and concentration, as requirements may vary by batch or supplier.
• For functional assays, confirm the biological activity and endotoxin level (<1 EU/µg is typical for cell culture).

Summary of key points:

• Reconstitute at 0.1 mg/mL in sterile water.
• Centrifuge vial before opening.
• Gently dissolve without vortexing.
• Aliquot and store appropriately.
• Minimize TFA exposure to cells by dilution or buffer exchange if necessary.

This protocol ensures optimal solubility and stability of recombinant chemerin for cell culture experiments.