Recombinant Human CXCL1

Using Recombinant Human CXCL1 in research applications enables precise, reproducible studies of CXCL1’s biological functions, including its roles in inflammation, immune cell recruitment, angiogenesis, tumor biology, and tissue remodeling.

Key scientific reasons to use recombinant human CXCL1:

• Defined, Consistent Activity: Recombinant CXCL1 provides a standardized, pure source of the chemokine, ensuring reproducibility and eliminating variability inherent in biological extracts.
• Functional Studies: It is essential for dissecting the specific effects of CXCL1 on target cells, such as:
  • Neutrophil chemotaxis: CXCL1 is a potent chemoattractant for neutrophils via CXCR2, making it valuable for migration and inflammation assays.
  • Angiogenesis: CXCL1 stimulates endothelial cell proliferation and tube formation, supporting studies on vascular biology and tumor angiogenesis.
  • Tumor Microenvironment: It modulates tumor growth, metastasis, and the recruitment of stromal and immune cells, relevant for cancer research.
  • Fibrosis and Tissue Remodeling: CXCL1 is implicated in wound healing, fibrosis, and tissue repair models.
• Disease Modeling: Recombinant CXCL1 is used to mimic or block its activity in models of cardiovascular, respiratory, and skin diseases, as well as in studies of infection and immune response.
• Assay Standardization: It serves as a standard in ELISA and other immunoassays for quantifying endogenous CXCL1 levels in biological samples.
• Mechanistic Dissection: Recombinant protein allows for controlled dose-response and mechanistic studies, including signaling pathway analysis (e.g., ERK1/2, EGF secretion).

Typical applications include:

• Cell migration and chemotaxis assays
• Angiogenesis and endothelial cell function assays
• Tumor cell proliferation and invasion studies
• ELISA and immunoassay standards
• In vitro and in vivo disease modeling

Summary:
Recombinant human CXCL1 is a critical tool for elucidating the chemokine’s roles in immunity, cancer, tissue repair, and inflammation, providing the specificity and consistency required for rigorous experimental design and interpretation.

Yes, you can use Recombinant Human CXCL1 (GROα) as a standard for quantification or calibration in your ELISA assays, provided that the recombinant protein is highly purified and specifically validated for use as an immunoassay standard.

Several sources confirm that recombinant human CXCL1 is routinely used as a standard in ELISA kits for generating standard curves and calibrating samples. For example:

• The R&D Systems Quantikine® Human CXCL1/GROα ELISA kit uses E. coli-expressed recombinant human GROα as its standard, and the assay is calibrated against this recombinant protein. The kit documentation states that the recombinant standard accurately quantitates both recombinant and natural human GROα, with results showing linear curves parallel to those obtained with the kit’s standards .
• Other ELISA kits, such as those from Thermo Fisher and Abcam, also specify the use of recombinant human CXCL1 as a standard for generating calibration curves [2, 4].
• The R&D Systems recombinant human CXCL1 protein (catalog 275-GR) is explicitly recommended for use as an immunoassay standard in ELISA .

Best Practices:

• Ensure the recombinant CXCL1 is lyophilized or supplied in a form suitable for reconstitution and dilution.
• Reconstitute and dilute the standard according to the manufacturer’s instructions or your assay protocol.
• Use the same diluent for both the standard and your samples to maintain consistency.
• Validate the standard’s performance in your specific assay by confirming linearity and parallelism with your samples.

In summary, recombinant human CXCL1 is a well-established and reliable standard for ELISA-based quantification and calibration of human CXCL1.

Recombinant Human CXCL1 has been validated for a range of applications in published research, primarily in the context of cell-based assays, immunoassays, and as a biomarker standard.

Key validated applications include:

• Bioassays (Cell-based functional assays):

  • Used to study neutrophil migration and activation, chemotaxis, and cell signaling pathways in various cell types, including endothelial cells, adipose stromal cells, cancer cells, and immune cells.
  • Applied in research on angiogenesis, chondrocyte differentiation, and tumor microenvironment modulation.
  • Employed to stimulate or modulate cellular responses in studies of osteoclastogenesis, senescence of cancer-associated fibroblasts, and leukocyte-endothelial adhesion.

• Immunoassays (ELISA):

  • Used as a standard protein for quantification of CXCL1 in biological samples, such as serum or cell culture supernatants.
  • Validated in studies measuring CXCL1 as a biomarker in clinical and preclinical samples, including in the context of acute respiratory distress syndrome and tuberculosis diagnosis.

• Diagnostic Biomarker Validation:

  • Validated as a serum biomarker for active tuberculosis (TB), meeting World Health Organization criteria for triage diagnostic tests with high sensitivity and specificity in human clinical samples.
  • Used in multiplex biomarker panels for disease discrimination (e.g., distinguishing active TB from latent TB infection and non-TB lung disease).

• Protein Characterization and Quality Control:

  • Validated by SDS-PAGE, HPLC, and mass spectrometry for purity and identity confirmation.

• Chemotaxis Assays:

  • Used to induce chemotaxis in transfected cell lines expressing CXCR2, with dose-dependent activity.

Summary Table of Validated Applications

Additional Notes:

• Recombinant Human CXCL1 is widely used in studies of inflammation, cancer biology, infectious disease, and tissue regeneration.
• It is important to select the appropriate formulation (e.g., carrier-free for cell culture, BSA-containing for ELISA standards) based on the intended application.

If you require protocols or more detailed application notes for a specific experimental context, please specify the intended use.

To reconstitute and prepare Recombinant Human CXCL1 protein for cell culture experiments, dissolve the lyophilized protein in sterile phosphate-buffered saline (PBS) or another appropriate sterile buffer to a concentration of 50–100 μg/mL. Use sterile technique throughout, and avoid vigorous pipetting or vortexing to prevent foaming and protein denaturation.

Step-by-step protocol:

• Bring the vial to room temperature before opening to minimize condensation.
• Add sterile PBS (or other recommended buffer) directly to the vial to achieve the desired concentration (commonly 100 μg/mL for stock solutions).
• Gently swirl or invert the vial to mix. Do not vortex.
• Allow the protein to fully dissolve (typically 10–30 minutes at room temperature). If necessary, pipette gently up and down to aid dissolution.
• Aliquot the reconstituted protein into single-use volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Store aliquots at –20 °C or –80 °C for long-term storage. Avoid repeated freeze-thaw cycles.

For cell culture use:

• Dilute the stock solution into your cell culture medium to the desired working concentration immediately before use. Use the same medium as your cells to avoid precipitation or osmotic shock.
• If the protein was lyophilized from a solution containing acetonitrile or trifluoroacetic acid (TFA), ensure complete removal of volatile solvents before use, or perform a buffer exchange if necessary.

Additional best practices:

• Always use sterile, low-protein-binding tubes and pipette tips.
• If using for sensitive cell types, consider adding a small amount of carrier protein (e.g., 0.1% BSA) to minimize adsorption to plastic, unless carrier-free conditions are required.
• Prepare fresh dilutions for each experiment and discard unused diluted protein.

Summary Table:

These steps ensure protein stability and biological activity for cell culture applications. Always consult the specific product datasheet for any lot-specific recommendations.