Recombinant Rat CXCL1/KC

Recombinant Rat CXCL1/KC is widely used in research because it is a key chemokine involved in neutrophil recruitment, inflammation, and immune response modulation in rat models. Using the recombinant form allows for controlled, reproducible studies of its biological effects in vitro and in vivo.

Key reasons to use recombinant Rat CXCL1/KC in research applications:

• Neutrophil Chemotaxis and Activation: CXCL1/KC is a potent chemoattractant for neutrophils, mediating their migration and activation during inflammatory responses. This makes it essential for studying mechanisms of acute and chronic inflammation, infection, and tissue injury in rat models.
• Modeling Disease Pathogenesis: Recombinant CXCL1/KC is used to mimic or modulate inflammatory conditions, such as autoimmune diseases, neuroinflammation, and demyelinating diseases, by controlling neutrophil infiltration and cytokine signaling.
• Functional Assays: The recombinant protein is biologically active and suitable for chemotaxis assays, receptor binding studies, and signaling pathway analyses, enabling precise investigation of CXCL1-mediated cellular responses.
• Reproducibility and Standardization: Recombinant CXCL1/KC provides a consistent, defined reagent compared to endogenous sources, reducing variability in experimental outcomes and facilitating quantitative studies.
• Mechanistic Studies: It allows for the dissection of CXCL1-specific effects on target cells (e.g., neutrophils, macrophages, epithelial cells) and the evaluation of downstream signaling pathways, such as those involving CXCR2, MAPK, and PI3K.

Additional applications include:

• Evaluating the role of CXCL1 in wound healing, cardiovascular, respiratory, and neurological diseases.
• Testing inhibitors or antibodies targeting CXCL1 or its receptor CXCR2 for therapeutic research.

In summary, recombinant Rat CXCL1/KC is a critical tool for dissecting the molecular and cellular mechanisms of inflammation and immune regulation in rat-based experimental systems, supporting both basic and translational research.

Yes, recombinant rat CXCL1/KC can be used as a standard for quantification and calibration in ELISA assays. This is a well-established practice in protein quantification workflows.

Use as ELISA Standards

Recombinant rat CXCL1 (also known as CINC-1 or KC/GRO-alpha) is specifically designed and validated for use as a calibration standard in sandwich ELISA assays. The recombinant protein is typically expressed in bacterial systems (such as E. coli) and serves as the reference material for generating standard curves.

Standard Curve Development

When using recombinant CXCL1 as your standard, the protein is reconstituted in an appropriate diluent (typically PBS or a reagent diluent containing BSA) and serially diluted to create a concentration range that brackets your expected sample values. The standard curve generated from these known concentrations allows you to interpolate the concentration of CXCL1 in your experimental samples by comparing their signal intensity to the standard response.

Key Considerations for Optimal Performance

Storage and Handling: Recombinant standards should be stored at -20°C or -80°C to maintain stability and activity. Importantly, you should avoid repeated freeze-thaw cycles, as these can compromise the protein's integrity and affect calibration accuracy.

Specificity: The recombinant standard will be recognized by the capture and detection antibody pairs in your sandwich ELISA, just as natural rat CXCL1 would be. This ensures that your calibration curve accurately reflects the assay's response to the target protein in your samples.

Lot-to-Lot Consistency: Quality-controlled recombinant standards are normalized across manufacturing lots to maintain consistent unit definitions and ensure reproducible calibration from assay to assay.

Using properly validated recombinant CXCL1 standards is therefore a reliable approach for accurate quantification of this chemokine in your ELISA experiments.

Recombinant rat CXCL1/KC has been validated for several important applications in published research, spanning both basic immunology and translational studies.

Neutrophil Biology and Host Defense

The primary validated application involves neutrophil recruitment and activation. Recombinant CXCL1 functions as a potent neutrophil attractant and activator, binding with high affinity to IL-8 receptor type B (CXCR2). This property has been extensively utilized in studies examining neutrophil-dependent immune responses, particularly in the context of bacterial lung infections. Research has demonstrated that recombinant CXCL1 can rescue impaired neutrophil-dependent host defense in knockout models, effectively restoring neutrophil mobilization from bone marrow and enhancing bacterial clearance in pulmonary infections.

Inflammatory and Immunological Studies

Recombinant rat CXCL1 has been validated for use in inflammation research, where it serves as a model chemokine for studying CXC chemokine-mediated immune responses. The protein exhibits chemotactic activity for neutrophils and plays documented roles in inflammatory cascade regulation.

Bioassay and Detection Applications

The protein has been validated for use in cell proliferation assays and in vitro bioactivity studies. Additionally, recombinant CXCL1 is employed in quantitative detection assays such as ELISA and multiplex immunoassays for measuring endogenous CXCL1 levels in biological samples including cell culture supernatants, serum, plasma, and urine.

Additional Biological Roles

Beyond neutrophil-focused applications, recombinant CXCL1 has been validated in studies examining angiogenesis, wound healing, and bone physiology, though these applications appear less extensively documented in the provided literature.

To reconstitute and prepare Recombinant Rat CXCL1/KC protein for cell culture experiments, first briefly centrifuge the vial to collect all lyophilized material at the bottom. Reconstitute the protein in sterile distilled water or sterile PBS to a concentration between 0.1–1.0 mg/mL. For optimal stability and to prevent adsorption, include 0.1% carrier protein such as bovine serum albumin (BSA) in the buffer.

Step-by-step protocol:

• Centrifuge the vial for 20–30 seconds in a microcentrifuge to ensure all powder is at the bottom.
• Add sterile distilled water or sterile PBS (with 0.1% BSA) to achieve the desired concentration (e.g., 100 μg/mL or up to 1 mg/mL).
• Gently mix by pipetting; do not vortex to avoid protein denaturation.
• Allow the protein to dissolve completely at room temperature for several minutes.
• If necessary, confirm reconstitution by SDS-PAGE (a band at ~7.8 kDa should be visible with as little as 10 ng loaded).

Storage after reconstitution:

• For short-term use (up to 1 week), store the solution at 2–8°C.
• For long-term storage, aliquot the solution (with carrier protein) and freeze at –20°C to –80°C.
• Avoid repeated freeze-thaw cycles to maintain protein integrity.

Preparation for cell culture:

• Thaw aliquots on ice and dilute to working concentrations in cell culture medium immediately before use.
• Ensure the final buffer is compatible with your cell culture system and does not exceed recommended carrier protein concentrations.

Additional notes:

• Always use sterile technique to prevent contamination.
• The presence of carrier protein (BSA) is critical for low concentration preparations (<0.5 mg/mL) to minimize loss due to adsorption.
• Do not vortex the protein solution at any stage.

This protocol ensures the recombinant CXCL1/KC protein remains bioactive and suitable for cell culture applications.