Recombinant Rat CXCL3

Recombinant Rat CXCL3 (C-X-C motif chemokine 3) is a valuable tool for research applications due to its well-characterized biological activity and its role in key cellular processes. Here are several reasons why you should consider using Recombinant Rat CXCL3 in your research:

1. Biological Relevance and Activity

• Recombinant Rat CXCL3 is fully biologically active and can be used to study chemokine-mediated signaling pathways. It is a potent neutrophil attractant and activator, similar to other GRO proteins, and its functional receptor is CXCR2.
• It is suitable for chemotaxis bioassays and other functional studies, allowing researchers to investigate the effects of CXCL3 on cell migration, proliferation, and activation.

2. Mechanistic Studies

• CXCL3 plays a significant role in tumorigenesis, inflammation, and immune responses. Using recombinant CXCL3 enables researchers to explore its effects on tumor cell differentiation, invasion, and migration, as well as its impact on gene expression (e.g., ERK1/2, Bcl-2, Bax, STAT3, NF-κB).
• It can be used to study the regulatory networks and signaling pathways associated with CXCL3, providing insights into its potential as a therapeutic target.

3. Versatility in Experimental Models

• Recombinant Rat CXCL3 can be used in both in vitro and in vivo studies. It is effective in cell culture systems for investigating cell proliferation, migration, and invasion, and can also be administered in animal models to study its effects on disease progression and immune responses.

4. Standardization and Reproducibility

• Using recombinant proteins ensures consistency and reproducibility in experiments. Recombinant Rat CXCL3 is available in standardized forms, which helps in obtaining reliable and comparable results across different studies.

5. Support for Drug Development and Therapeutic Research

• Understanding the role of CXCL3 in various diseases, such as cancer and inflammatory conditions, can inform the development of new therapeutic strategies. Recombinant CXCL3 can be used to screen for inhibitors or modulators of CXCL3 signaling, contributing to drug discovery efforts.

6. Compatibility with Various Assays

• Recombinant Rat CXCL3 is compatible with a wide range of assays, including ELISA, Western blotting, and functional assays like chemotaxis and cell proliferation assays. This versatility makes it a useful reagent for diverse research applications.

In summary, Recombinant Rat CXCL3 is a powerful tool for studying the biological functions of CXCL3, its role in disease, and its potential as a therapeutic target. Its biological activity, versatility, and compatibility with various experimental models make it an essential reagent for researchers in the fields of immunology, oncology, and inflammation.

Yes, you can use recombinant rat CXCL3 as a standard for quantification or calibration in your ELISA assays, provided it is of high purity and its concentration is accurately known. This is a common and accepted practice in quantitative ELISA protocols.

Key considerations and supporting details:

• Recombinant proteins are routinely used as ELISA standards: Quantitative ELISA assays require a standard curve generated from known concentrations of the target protein. Recombinant CXCL3, if properly quantified and pure, is suitable for this purpose.
• Commercial ELISA kits often use recombinant CXCL3 as the standard: Many rat CXCL3 ELISA kits specify that their standards are recombinant proteins, and these are validated for generating standard curves that allow accurate quantification of CXCL3 in biological samples.
• Purity and quantification are critical: The recombinant CXCL3 should be highly purified, and its concentration must be determined accurately (e.g., by absorbance at 280 nm, BCA assay, or HPLC). Impurities or inaccurate quantification can lead to errors in your standard curve and sample measurements.
• Matrix compatibility: If your samples are in a complex matrix (e.g., serum, plasma), ensure that the recombinant standard is diluted in a similar matrix or in the assay buffer recommended by your ELISA protocol to minimize matrix effects.
• Carrier proteins: Some recombinant proteins are supplied with carrier proteins (e.g., BSA) to enhance stability. If your recombinant CXCL3 contains a carrier, ensure this is compatible with your assay and does not interfere with antibody binding.
• Validation: If you are developing a custom ELISA or using a recombinant standard not provided with a commercial kit, it is good practice to validate the standard curve by checking for parallelism with endogenous CXCL3 in your sample matrix.

Protocol summary for using recombinant CXCL3 as an ELISA standard:

• Reconstitute the recombinant protein according to the manufacturer’s instructions or your lab’s standard protocol.
• Prepare a series of serial dilutions covering the expected range of CXCL3 concentrations in your samples.
• Run these standards in parallel with your samples to generate a standard curve.
• Use the standard curve to interpolate the concentrations of CXCL3 in your unknown samples.

In summary: Recombinant rat CXCL3 is appropriate for use as a standard in ELISA quantification, provided it is pure, accurately quantified, and compatible with your assay conditions. Always follow best practices for standard preparation and validation to ensure reliable results.

Recombinant Rat CXCL3 has been validated in published research primarily for applications related to its chemotactic and inflammatory properties, especially in the context of neutrophil recruitment, inflammation, and cancer biology.

Key validated applications include:

• In vitro chemotaxis assays: Recombinant rat CXCL3 is widely used to assess its ability to induce migration of neutrophils and other immune cells, confirming its chemotactic activity via CXCR2.
• Cell signaling studies: It is used to stimulate endothelial cells and other target cells to study downstream signaling pathways, such as activation of MAPK/ERK and PI3K/Akt, and to investigate autocrine effects in inflammation.
• Inflammation models: Recombinant CXCL3 is applied in cell culture and animal models to study its role in inflammatory responses, including wound healing and tissue injury.
• Cancer research: It has been used to investigate the role of CXCL3 in tumor progression, angiogenesis, and metastasis, particularly in models of medulloblastoma and colorectal cancer.
• ELISA standard and positive control: Recombinant CXCL3 is validated as a standard in ELISA assays for quantifying endogenous CXCL3 levels in biological samples.

Supporting details:

• Chemotaxis and cell activation: Multiple studies confirm that recombinant rat CXCL3 induces chemotaxis of neutrophils and activates inflammatory pathways, validating its use in migration and activation assays.
• Endothelial and immune cell assays: It is used to probe the effects of CXCL3 on endothelial cells, including autocrine signaling and modulation of inflammatory gene expression.
• Cancer models: Recombinant CXCL3 has been used to manipulate tumor microenvironments, assess forced migration of tumor cells, and evaluate therapeutic potential in preclinical cancer models.
• ELISA and functional assays: Commercial and academic protocols validate recombinant CXCL3 as a standard for ELISA and as a positive control in functional cell-based assays.

Summary of best practices:

• Use carrier-free recombinant CXCL3 for cell-based functional assays to avoid interference from stabilizing proteins.
• Validate biological activity in each experimental system, as chemokine effects can be context-dependent.

No evidence was found for use in diagnostic or therapeutic applications in humans; all validated uses are for research purposes only.

To reconstitute and prepare Recombinant Rat CXCL3 protein for cell culture experiments, dissolve the lyophilized protein in sterile PBS containing at least 0.1% human or bovine serum albumin (BSA) to a final concentration of 50 μg/mL. This carrier protein helps stabilize CXCL3 and prevents adsorption to tube walls, which is critical for bioactivity in cell culture assays.

Step-by-step protocol:

• Centrifuge the vial briefly to collect the powder at the bottom before opening.
• Add sterile PBS + 0.1% BSA to achieve the desired concentration (e.g., 50 μg/mL).
• Gently mix by pipetting up and down or by slow vortexing until fully dissolved. Avoid vigorous agitation to prevent protein denaturation.
• Aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Store aliquots at 4°C for up to 1 week, or at −20°C for up to 2 months.

Additional notes for cell culture use:

• If your protocol or supplier recommends a different buffer (e.g., sterile water or other diluents), always check the product datasheet for specific instructions.
• For most cell-based assays, working concentrations of CXCL3 typically range from 10–100 ng/mL; dilute the stock solution in your cell culture medium immediately before use.
• If using serum-free medium, ensure the presence of carrier protein (BSA or similar) to maintain CXCL3 stability.
• Avoid multiple freeze-thaw cycles and prolonged storage at room temperature to preserve bioactivity.

Summary Table: Recombinant Rat CXCL3 Reconstitution

Always consult the specific product datasheet for any unique requirements, and ensure all solutions are sterile to prevent contamination in cell culture experiments.