Recombinant Human CXCL10

Recombinant Human CXCL10 (also known as IP-10) is a valuable tool for a wide range of research applications due to its well-characterized biological activities and roles in immune regulation, inflammation, and host defense. Here are several key reasons why you should consider using Recombinant Human CXCL10 in your research:

1. Immune Cell Recruitment and Activation

CXCL10 is a potent chemoattractant for activated T cells (especially Th1 cells), natural killer (NK) cells, dendritic cells, monocytes, and macrophages. By using recombinant CXCL10, you can:

• Study the migration and recruitment of immune cells in vitro or in vivo.
• Investigate the mechanisms of immune surveillance and inflammatory responses.

2. Regulation of Adaptive and Innate Immunity

CXCL10 plays a critical role in bridging innate and adaptive immunity by:

• Recruiting and activating T cells and NK cells at sites of infection or inflammation.
• Modulating dendritic cell and macrophage function.

3. Antiviral and Antitumor Effects

CXCL10 is upregulated in response to viral infections and has been shown to:

• Enhance antiviral immune responses.
• Exert anti-proliferative and angiostatic effects on endothelial cells, contributing to tumor suppression.

4. Antimicrobial Activity

Recent studies have demonstrated that CXCL10 possesses direct antimicrobial activity against a broad spectrum of bacteria, including:

• Escherichia coli
• Listeria monocytogenes
• Bacillus anthracis
• Staphylococcus aureus
• Carbapenem-resistant and other multidrug-resistant pathogens

This makes recombinant CXCL10 a useful molecule for studying host defense mechanisms and the development of novel antimicrobial strategies.

5. Inflammatory Disease Models

CXCL10 is strongly upregulated in many inflammatory and autoimmune diseases, such as:

• Rheumatoid arthritis
• Type I diabetes
• Multiple sclerosis
• Atherosclerosis
• Allograft rejection

Using recombinant CXCL10 allows you to model and study these conditions, as well as investigate the role of chemokines in disease pathogenesis.

6. Signal Transduction Studies

CXCL10 binds to the CXCR3 receptor, activating G protein-mediated signaling pathways. This leads to:

• Activation of phospholipase C
• Increase in intracellular calcium
• Actin reorganization

These effects make CXCL10 ideal for studying chemokine receptor signaling and downstream cellular responses.

7. Standard for Assays

Recombinant CXCL10 is commonly used as a standard in:

• ELISA assays
• Bioassays
• Functional studies

This ensures accurate quantification and reproducibility in your experiments.

8. Therapeutic Development

The identification of distinct antimicrobial domains within CXCL10 opens up possibilities for developing novel peptide-based therapeutics targeting antibiotic-resistant bacteria.

In summary, Recombinant Human CXCL10 is a versatile and powerful reagent for studying immune responses, inflammation, host defense, and disease mechanisms. Its broad range of biological activities makes it an essential tool for researchers in immunology, infectious diseases, cancer biology, and related fields.

Yes, recombinant human CXCL10 can be used as a standard for quantification or calibration in ELISA assays, provided that the ELISA kit is designed to detect human CXCL10 and the recombinant protein is of high purity and appropriate for immunoassay use.

Several ELISA kits for human CXCL10 (IP-10) specifically mention that they can accurately quantitate both natural and recombinant human CXCL10. For example:

• R&D Systems' Quantikine and DuoSet ELISA kits state that they recognize both natural and recombinant human CXCL10 and can be used to determine relative mass values for natural human CXCL10 using recombinant standards.
• Cytodiagnostics, Abcam, Sigma-Aldrich, and other manufacturers also indicate that their kits are validated for use with recombinant human CXCL10 standards.

When using recombinant human CXCL10 as a standard, ensure that:

• The protein is properly reconstituted and diluted according to the manufacturer's instructions.
• The standard curve is prepared fresh for each assay.
• The recombinant protein is compatible with the detection antibodies and assay buffer system used in your ELISA kit.

In summary, recombinant human CXCL10 is suitable for use as a standard in ELISA assays for quantifying human CXCL10, as long as the ELISA kit is validated for this purpose and the recombinant protein is of high quality.

Recombinant Human CXCL10 has been validated for a range of applications in published research, primarily in studies of immunology, inflammation, cancer, and infectious disease. The most commonly validated applications include:

• Bioassays / Functional Assays: Used to study chemotactic activity, immune cell recruitment (e.g., T cells, NK cells, dendritic cells), and signaling via the CXCR3 receptor in vitro and in vivo.
• ELISA (Standard): Utilized as a standard for quantifying CXCL10 levels in biological samples, such as serum or cell culture supernatants, in studies of disease biomarkers and immune responses.
• Western Blot: Applied for protein detection and validation in cell lysates and tissue samples.
• Protein-Protein Interaction / Binding Assays: Used to investigate interactions with receptors (CXCR3) and other proteins.
• Antimicrobial Activity Assays: CXCL10 and its derived peptides have been validated for bactericidal activity against a broad spectrum of pathogens, including antibiotic-resistant bacteria, in vitro.
• Cell Culture / Immunotherapy Models: Incorporated into cell culture systems to study effects on cell migration, adhesion, and immune modulation, including engineered fusion proteins for enhanced NK cell recruitment.
• In Vivo Models: Used in animal studies to investigate CXCL10’s role in inflammation, infection, and immune cell trafficking.

Supporting Details and Examples:

• Chemotaxis and Immune Cell Recruitment: CXCL10 is frequently used to induce migration of CXCR3-expressing cells (e.g., Th1 lymphocytes, NK cells, dendritic cells) in transwell assays and flow-based adhesion models.
• Disease Biomarker Studies: CXCL10 is validated as a biomarker in ELISA-based studies for conditions such as lymphoma, cervical cancer, and autoimmune diseases.
• Antimicrobial Mechanisms: Research has mapped bactericidal domains within CXCL10, demonstrating direct antimicrobial effects against Gram-positive and Gram-negative bacteria, including multidrug-resistant strains.
• Cell Signaling: Functional assays confirm CXCL10’s ability to activate CXCR3-mediated G protein signaling, leading to downstream effects such as calcium flux and actin reorganization.
• Protein Characterization: SDS-PAGE, HPLC, and mass spectrometry are used for purity and identity confirmation.

Summary Table of Validated Applications

These applications are supported by multiple peer-reviewed publications and product validation data, demonstrating the versatility of recombinant human CXCL10 in immunological and translational research.

To reconstitute and prepare Recombinant Human CXCL10 protein for cell culture experiments, dissolve the lyophilized protein in sterile PBS or water to a concentration of 100 μg/mL. If the formulation includes carrier protein (such as BSA), use PBS containing at least 0.1% human or bovine serum albumin to enhance stability and prevent adsorption to surfaces.

Step-by-step protocol:

• Briefly centrifuge the vial to collect the powder at the bottom before opening.
• Add sterile PBS (or water, if recommended by the manufacturer) to achieve the desired concentration, typically 100 μg/mL.
• If the protein is supplied without carrier, use plain sterile PBS. If carrier is present or recommended, add at least 0.1% BSA to the PBS.
• Gently mix by inverting or flicking the tube. Avoid vortexing, as vigorous mixing can denature the protein.
• Wait 5 minutes to allow complete dissolution, then gently mix again.
• Aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Store aliquots at −20°C or −70°C for long-term storage. For short-term use (2–7 days), store at 4–8°C.
• Before use in cell culture, dilute the stock solution to the working concentration using cell culture medium or PBS with 0.1% BSA, as appropriate for your assay.

Additional notes:

• For bioactivity assays, typical working concentrations range from 0.03–0.18 μg/mL.
• Always check the specific product datasheet for recommended reconstitution buffer and concentration, as formulations may vary.
• Avoid repeated freeze-thaw cycles to maintain protein integrity.
• If using for ELISA or other analytical assays, follow the recommended diluent and protocol for those applications.

This protocol ensures optimal solubility and stability of recombinant CXCL10 for cell culture and functional assays.