Recombinant Mouse Lungkine

Recombinant Mouse Lungkine (CXCL15) is a chemokine with distinct biological activities, making it valuable for research applications involving hematopoiesis, immune cell migration, and lung biology. Its use is justified when investigating the regulation of hematopoietic progenitor cell (HPC) proliferation, neutrophil trafficking, or pulmonary immune responses.

Key scientific reasons to use Recombinant Mouse Lungkine:

• Negative Regulation of Hematopoietic Progenitor Cell Cycling:
  Recombinant Lungkine suppresses the proliferation of bone marrow and spleen HPCs (including CFU-GM, BFU-E, and CFU-GEMM) during cytokine-enhanced ex vivo culture. This myelosuppressive effect is S-phase specific and can be used to study mechanisms of HPC regulation, cell cycle control, and myelosuppression in vitro.

• Mediator of Neutrophil Migration:
  Lungkine is a critical chemokine for neutrophil recruitment from lung parenchyma into the airspace during pulmonary inflammation. Recombinant Lungkine can be used to model or manipulate neutrophil trafficking in studies of lung injury, infection, or inflammation.

• Functional Studies in Knockout and Wild-Type Models:
  Using recombinant Lungkine allows for direct comparison of its effects in wild-type versus CXCL15/Lungkine knockout mice, helping to dissect its physiological roles in hematopoiesis and immune cell migration.

• Molecular and Cellular Mechanism Elucidation:
  Recombinant protein enables controlled, dose-dependent studies of Lungkine’s effects on cell signaling, gene expression, and functional outcomes in primary cells or cell lines.

Typical research applications include:

• Ex vivo culture of mouse bone marrow or spleen cells to assess HPC proliferation and colony formation under cytokine stimulation.
• In vitro assays of neutrophil chemotaxis and migration, especially in pulmonary models.
• Studies of lung inflammation, infection, or injury, where neutrophil recruitment is a key endpoint.
• Mechanistic investigations into chemokine signaling pathways and their impact on hematopoietic or immune cell function.

Technical considerations:

• Recombinant Mouse Lungkine is typically supplied as a non-glycosylated polypeptide of ~16–18 kDa, suitable for cell culture, migration assays, and biochemical studies.
• Dose and timing should be optimized based on the specific cell type and experimental design.

In summary:
Use Recombinant Mouse Lungkine when you need to experimentally modulate or study the suppression of hematopoietic progenitor proliferation, neutrophil migration in lung models, or the molecular mechanisms of CXCL15/Lungkine signaling in mouse systems.

You can use recombinant mouse Lungkine (CXCL15) as a standard for quantification or calibration in your ELISA assays, provided it is of high purity and its concentration is accurately known. Recombinant proteins are commonly used as standards in ELISA to generate a standard curve, which is essential for quantifying the analyte in unknown samples.

Key considerations:

• Purity and Quantification: The recombinant Lungkine should be highly purified, and its concentration should be determined precisely, ideally by methods such as HPLC or absorbance at 280 nm with a known extinction coefficient.
• Standard Curve Preparation: Prepare a serial dilution of the recombinant protein in the same buffer or matrix as your samples to generate a standard curve for quantification.
• Assay Compatibility: Ensure that the antibodies used in your ELISA recognize both the recombinant and native forms of Lungkine equivalently. Most commercial ELISA kits for mouse CXCL15/Lungkine are validated to detect both natural and recombinant forms, and their protocols include recombinant Lungkine as the standard.
• Matrix Effects: If you are quantifying Lungkine in complex matrices (e.g., serum, plasma), validate that the recombinant standard behaves similarly to the native protein in your assay conditions.

Protocol Example:

• Coat the ELISA plate with capture antibody.
• Block, then add serial dilutions of recombinant Lungkine to generate the standard curve.
• Add samples, detection antibody, and develop as per your assay protocol.

Summary Table:

References to 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".
• "This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse CXCL15/Lungkine".
• "In order for the assay to be quantitative, a standard curve must be prepared for each assay".

Conclusion:
Recombinant mouse Lungkine is suitable as a standard for ELISA quantification, provided it is pure, accurately quantified, and validated for use with your assay antibodies and sample matrix.

Recombinant Mouse Lungkine (CXCL15) has been validated in published research for applications including in vitro colony formation assays, ex vivo hematopoietic progenitor cell culture, and studies of neutrophil migration and myelosuppression.

Key validated applications in the literature:

• In vitro colony formation assays: Recombinant Mouse Lungkine has been used to suppress colony formation of hematopoietic progenitor cells (HPCs) from wildtype mouse bone marrow, specifically CFU-GM, CFU-G, CFU-M, BFU-E, and CFU-GEMM, demonstrating its myelosuppressive activity.

• Ex vivo culture of hematopoietic progenitors: It has been applied to lineage-negative mouse bone marrow cells in multi-cytokine stimulated cultures to assess its regulatory effects on HPC proliferation and cycling.

• Neutrophil migration studies: Recombinant Lungkine has been used to investigate its role as a mediator of neutrophil trafficking from lung parenchyma into the airspace, particularly in models of pulmonary inflammation.

• Immunological and cell signaling research: Its effects on cell cycling and myelosuppression have been explored in knockout and wildtype mouse models to elucidate its physiological role in hematopoiesis and immune cell regulation.

• Detection and quantification assays: Recombinant Mouse Lungkine is used as a standard or control in ELISA and Western blot assays for the detection of endogenous CXCL15/Lungkine in biological samples.

Summary Table of Validated Applications

Additional Notes:

• Recombinant Mouse Lungkine has not been widely reported in organoid or cancer model applications, nor in skin graft or tissue regeneration studies, based on current published research.
• Most functional studies focus on its role in hematopoiesis and immune cell migration, particularly in the context of lung and bone marrow biology.

If you require protocols or further details for a specific application, please specify the experimental context.

To reconstitute and prepare Recombinant Mouse Lungkine protein for cell culture experiments, first briefly centrifuge the vial to collect all lyophilized material at the bottom, then add sterile deionized water or an appropriate buffer to achieve a concentration between 0.1–1.0 mg/mL, optionally including 0.1%–1% BSA as a carrier protein to enhance stability.

Detailed protocol:

• Centrifuge the vial briefly before opening to ensure all powder is at the bottom.
• Reconstitution:
  • Add sterile deionized water or sterile PBS to the vial. Commonly recommended concentrations are:
    • 0.1–1.0 mg/mL for general use.
    • Some protocols recommend at least 100 µg/mL as a minimum concentration.
    • If using PBS, include at least 0.1% BSA (bovine serum albumin) to prevent protein adsorption and enhance stability, especially for storage or low-concentration use.
• Mix gently by swirling or pipetting up and down. Do not vortex or create foam, as this can denature the protein.
• Allow the solution to stand at room temperature for 1–15 minutes to ensure complete dissolution.
• Aliquot the solution into single-use volumes to avoid repeated freeze-thaw cycles, which can reduce protein activity.
• Storage:
  • Store aliquots at -20°C to -80°C for long-term use.
  • For short-term use (up to 1 week), store at 2–8°C.
  • If storing at concentrations below 50 µg/mL, always add a carrier protein such as 1% BSA to prevent loss of activity.
• Avoid repeated freeze-thaw cycles to maintain protein integrity.

Additional notes:

• Always use sterile technique to prevent contamination.
• If the protein does not fully dissolve, gentle mixing for up to 2 hours at room temperature may be necessary.
• Check the specific product datasheet for any unique requirements, as some preparations may require acidic buffers (e.g., 10 mM HCl) to prevent precipitation, though for Lungkine, water or PBS with BSA is standard.

Summary Table:

This protocol ensures optimal solubility, stability, and bioactivity of recombinant Mouse Lungkine for cell culture applications.