Recombinant Mouse CXCL2

Recombinant Mouse CXCL2 is widely used in research applications to study immune cell migration, inflammation, and tumor microenvironment dynamics due to its potent ability to chemoattract and activate neutrophils. Using recombinant CXCL2 allows for controlled, reproducible experiments investigating the molecular mechanisms of neutrophil recruitment and related immune responses.

Key scientific reasons to use recombinant Mouse CXCL2 include:

• Neutrophil Chemotaxis and Activation: CXCL2 is a major chemokine responsible for attracting neutrophils to sites of inflammation or infection, making it essential for in vitro migration assays, transwell experiments, and in vivo studies of immune cell trafficking.
• Modeling Inflammatory Responses: CXCL2 is upregulated by pro-inflammatory cytokines (e.g., TNFα, IL-1α, IL-17, LPS) and is involved in the mobilization of neutrophils from the bone marrow, allowing researchers to model acute and chronic inflammatory conditions.
• Tumor Microenvironment Studies: CXCL2 plays a pivotal role in cancer biology by modulating immune cell infiltration, particularly neutrophils and myeloid cells, which can influence tumor progression and response to immunotherapies.
• Functional Assays: Recombinant CXCL2 is used in bioassays to study cell migration, adhesion, and activation, and is suitable for ELISA standardization, binding assays, and chemotaxis assays.
• Reproducibility and Purity: Recombinant proteins provide high purity and batch-to-batch consistency, which is critical for quantitative and mechanistic studies.

Typical applications include:

• In vitro neutrophil migration assays (e.g., transwell, random migration, adhesion assays).
• In vivo studies of neutrophil recruitment and inflammation.
• Investigating the role of CXCL2 in disease models such as sepsis, autoimmune disorders, and cancer.
• ELISA development and standardization for CXCL2 quantification.

Best practices:
Use recombinant CXCL2 at concentrations validated for your assay type (often in the range of 0.1–100 ng/mL for chemotaxis assays). Ensure the protein is carrier-free if required for sensitive applications, and confirm bioactivity with appropriate controls.

In summary, recombinant Mouse CXCL2 is a critical tool for dissecting the mechanisms of neutrophil migration, inflammation, and immune regulation in mouse models, enabling precise and reproducible experimental outcomes.

Yes, recombinant Mouse CXCL2 (also known as MIP-2) can be used as a standard for quantification or calibration in ELISA assays, provided it is formulated and purified appropriately for this purpose.

Several sources confirm that recombinant Mouse CXCL2 is routinely used as a standard in ELISA kits and development systems:

• R&D Systems and BioLegend offer recombinant Mouse CXCL2 specifically for use as an ELISA standard, with formulations in PBS and high purity (>98% by SDS-PAGE).
• ELISA development kits (such as R&D Systems DuoSet DY452, Bio-Techne DY452, Thermo Fisher 900-K152K, and Leinco Technologies) include recombinant Mouse CXCL2 as the protein standard for generating standard curves.
• Abcam, Thermo Fisher, and Proteintech ELISA kits also use recombinant Mouse CXCL2 as the reference standard for quantifying endogenous CXCL2 in samples.

Important considerations:

• Ensure the recombinant protein is carrier-free (without BSA or other stabilizers) if you are using it as an ELISA standard, as carriers can interfere with quantification.
• Verify that the recombinant CXCL2 is purified and tested for endotoxin levels, as high endotoxin can affect assay performance.
• Always follow the manufacturer’s instructions for reconstitution and dilution to ensure accurate standard curve generation.

In summary, recombinant Mouse CXCL2 is suitable and commonly used as a standard for ELISA quantification, provided it is of high purity and appropriate formulation.

Recombinant Mouse CXCL2 has been validated for several key applications in published research, primarily involving its role as a chemotactic factor and immune modulator. The most frequently validated applications include:

• Bioassays (Functional Assays):

  • Used to assess neutrophil chemotaxis in vitro, often in transwell migration assays, where recombinant CXCL2 serves as a chemoattractant for mouse neutrophils.
  • Applied in studies of cell signaling and immune cell recruitment, such as investigating the mechanisms of neutrophil and macrophage migration in response to inflammatory stimuli.

• In Vivo Studies:

  • Employed to study leukocyte recruitment and inflammation in mouse models, including peritonitis, heart transplantation, and tumor microenvironment research.
  • Used to modulate immune responses and evaluate the effects of CXCL2 on disease progression, such as in models of arthritis, sepsis, and cancer metastasis.

• ELISA Development and Standardization:

  • Utilized as a standard protein for the development and calibration of ELISA assays to quantify endogenous CXCL2 levels in tissue homogenates, serum, plasma, and cell culture supernatants.

• Cell Migration/Motility Assays:

  • Validated as a positive control in in vitro chemotaxis assays, particularly for neutrophil migration, confirming its bioactivity and specificity in these systems.

• Other Functional Studies:

  • Used in studies examining the CXCL2-CXCR2 axis in cancer biology, angiogenesis, and immune cell infiltration, including mechanistic investigations of tumor growth and metastasis.

Summary Table of Validated Applications:

Key details:

• Recombinant Mouse CXCL2 is most commonly validated for bioassays and in vivo studies related to immune cell migration, especially neutrophils.
• It is also widely used as an ELISA standard and in cell migration assays.
• Published research supports its use in both basic immunology and disease models, including cancer, infection, and inflammatory diseases.

If you need protocols or more specific examples for any of these applications, please specify the context or experimental system.

To reconstitute and prepare Recombinant Mouse CXCL2 protein for cell culture experiments, dissolve the lyophilized protein at a concentration of 100 μg/mL in sterile PBS. For optimal stability and to minimize adsorption, it is recommended to include 0.1–1% carrier protein such as bovine serum albumin (BSA) or human serum albumin (HSA) in the PBS.

Detailed protocol:

• Reconstitution:

  • Briefly centrifuge the vial to collect the lyophilized powder at the bottom before opening.
  • Add sterile PBS (pH 7.2–7.4) to achieve a final concentration of 100 μg/mL.
  • If the protein is sensitive or will be used at low concentrations, add 0.1–1% BSA or HSA to the PBS to prevent loss due to adsorption.
  • Gently mix by pipetting up and down or by slow vortexing. Avoid vigorous agitation or foaming.

• Aliquoting and Storage:

  • Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can reduce activity.
  • Store aliquots at –20°C to –70°C for long-term storage. For short-term use (up to 1 week), store at 2–8°C.
  • Avoid multiple freeze-thaw cycles.

• Preparation for Cell Culture:

  • Before use, dilute the stock solution to the desired working concentration using sterile cell culture medium or PBS containing 0.1% BSA.
  • Typical working concentrations for chemotaxis or signaling assays range from 0.1–100 ng/mL, depending on the cell type and assay sensitivity.

Additional notes:

• If the product datasheet specifies a different buffer (e.g., sterile water or a specific buffer with stabilizers), follow those instructions.
• Always filter-sterilize the final working solution if sterility is required and the protein solution is not already sterile-filtered.
• Confirm the absence of endotoxin if using in sensitive cell culture systems; most recombinant preparations are tested to be <0.01 ng/μg.

This protocol ensures maximal recovery, stability, and biological activity of recombinant mouse CXCL2 for cell-based assays.