Recombinant Human Lymphotactin

Recombinant Human Lymphotactin (XCL1) is a valuable tool for a wide range of research applications due to its well-characterized biological activity and relevance in immunology, cell biology, and disease mechanisms. Here are several key reasons why you should consider using Recombinant Human Lymphotactin in your research:

1. Immunological Research

• Lymphotactin is a chemokine that specifically recruits T cells and natural killer (NK) cells, making it essential for studying immune cell migration, activation, and function.
• It plays a critical role in dendritic cell-mediated cytotoxic immune responses and the development of regulatory T cells, which are important for understanding immune tolerance and autoimmune diseases.

2. Cell Biology Studies

• Recombinant Lymphotactin can be used to investigate cell signaling pathways, protein-protein interactions, and the effects of chemokine stimulation on various cell types.
• It is useful for studying the mechanisms of chemotaxis and cell migration in both normal and pathological conditions.

3. Therapeutic Development

• Lymphotactin has been explored in cancer immunotherapies and continues to be a subject of ongoing research for its potential therapeutic applications.
• Its ability to modulate immune responses makes it a candidate for developing new treatments for cancer, autoimmune diseases, and other immune-related disorders.

4. Functional Assays

• Recombinant Lymphotactin is validated for use in various functional assays, including blocking assays, ELISA, and Western blot, allowing for comprehensive analysis of its biological effects.
• It can be used to study the interaction between Lymphotactin and its receptor, XCR1, and to evaluate the impact of this interaction on cellular processes.

5. Neurological Research

• Recent studies have identified a role for Lymphotactin (XCL1) signaling in nociceptive processing and neurological disorders such as multiple sclerosis, Parkinson’s, Huntington’s, and Alzheimer’s diseases.
• Using recombinant Lymphotactin can help elucidate the mechanisms underlying these conditions and identify potential therapeutic targets.

6. High Biological Activity

• Recombinant Lymphotactin is fully biologically active and can be used in chemotaxis bioassays to study the recruitment of lymphocytes and NK cells.
• The protein is typically expressed in E. coli and is available in various forms, including carrier-free and with tags, to suit different experimental needs.

7. Quality and Reliability

• Many suppliers provide recombinant Lymphotactin with rigorous quality control testing, ensuring high purity and activity.
• Products are often covered by guarantees, providing confidence in their performance and reliability.

8. Versatile Applications

• Recombinant Lymphotactin can be used in a variety of applications, including immunological research, cell biology studies, protein-protein interaction analyses, and therapeutic development.
• It is suitable for both in vitro and in vivo studies, making it a versatile tool for researchers in multiple fields.

By using Recombinant Human Lymphotactin, you can gain insights into the complex mechanisms of immune cell recruitment, signaling, and function, as well as explore its potential in therapeutic and diagnostic applications.

Yes, recombinant human lymphotactin (XCL1) can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately known. This is a common and accepted practice in cytokine and chemokine quantification assays.

Key considerations and supporting details:

• Recombinant proteins are routinely used as ELISA standards: Most commercial ELISA kits for human lymphotactin/XCL1 use recombinant human XCL1 as the standard to generate the calibration curve for quantification. The standard curve is essential for determining the concentration of XCL1 in unknown samples.

• Purity and formulation matter: For best results, use a recombinant protein that is highly purified and free of contaminants. Carrier-free formulations are preferred if BSA or other carriers could interfere with your assay, but BSA-containing formulations are also commonly used for ELISA standards to enhance stability.

• Preparation and handling: Follow the manufacturer’s instructions for reconstitution and dilution of the recombinant standard. Accurate pipetting and preparation of serial dilutions are critical for generating a reliable standard curve.

• Validation: Ensure that the recombinant standard is compatible with your ELISA system (e.g., antibody specificity, detection range). Most ELISA kits are validated to detect both natural and recombinant forms of the target protein.

• Concentration range: The standard curve should cover the expected concentration range of your samples. Typical ELISA standard curves for XCL1 range from low picogram to nanogram per milliliter concentrations.

Summary of best practices:

• Use a recombinant human lymphotactin/XCL1 of known concentration and high purity.
• Prepare standards according to validated protocols, using appropriate diluents and avoiding repeated freeze-thaw cycles.
• Confirm that your ELISA antibodies recognize the recombinant form used as the standard.
• Generate a standard curve in every assay run for accurate quantification.

In conclusion, recombinant human lymphotactin is suitable and widely used as a standard for ELISA quantification, provided these technical considerations are met.

Recombinant Human Lymphotactin (XCL1) has been validated in published research for a range of applications, including functional bioassays, chemotaxis assays, blocking assays, ELISA, Western blot, binding assays, and studies of antimicrobial and antiviral activity.

Key validated applications and research uses include:

• Chemotaxis and Functional Bioassays:
  XCL1 is widely used to assess chemotactic activity for lymphocytes, NK cells, and dendritic cells, often via migration or chemotaxis assays. Its ability to chemoattract BaF3 mouse pro B cells transfected with human XCR1 is a standard bioactivity assay.

• Blocking Assays:
  Recombinant XCL1 has been used in blocking assays to study its ability to inhibit specific cellular interactions or signaling pathways.

• ELISA and Western Blot:
  The protein is validated for use as a standard or control in ELISA and Western blot protocols, supporting detection and quantification of XCL1 or its receptor interactions.

• Binding Assays:
  XCL1 has been used in binding assays to characterize its interaction with receptors such as XCR1 and to study ligand-receptor specificity.

• Antimicrobial and Antifungal Activity:
  Published studies have demonstrated that XCL1 exhibits direct antimicrobial activity, including membrane disruption of bacterial and fungal cells (notably Candida species), independent of classical immune cell-mediated mechanisms. This is often assessed using in vitro killing or membrane permeabilization assays.

• Antiviral Activity:
  XCL1 has been shown to act as a conformation-dependent inhibitor of HIV-1, blocking viral attachment and entry into host cells. These effects are typically measured using viral entry or infectivity assays.

• Cellular and Molecular Immunology Studies:
  XCL1 is used to investigate its role in T cell and NK cell recruitment, dendritic cell-mediated cytotoxic responses, and the development of regulatory T cells.

• Bioassays for Growth Factor and Apoptosis Studies:
  Research has used XCL1 in bioassays to study its anti-apoptotic effects and its involvement in resistance to anticancer kinase inhibitors.

• Protein Characterization:
  SDS-PAGE is used for quality control and characterization of recombinant XCL1.

Summary Table of Validated Applications

These applications are supported by both product validation data and peer-reviewed research, demonstrating the versatility of recombinant human lymphotactin in immunological, microbiological, and biochemical studies.

To reconstitute and prepare Recombinant Human Lymphotactin (XCL1) for cell culture experiments, follow these steps for optimal protein stability and biological activity:

1. Preparation Before Reconstitution

• Briefly centrifuge the vial (3,000–3,500 rpm, 5 min) to collect all lyophilized powder at the bottom and avoid loss when opening.
• Allow both the vial and the reconstitution buffer to equilibrate to room temperature before proceeding.

2. Reconstitution

• For most lyophilized recombinant human lymphotactin proteins, reconstitute at 100 μg/mL in sterile PBS.
• If the formulation contains no carrier protein, add at least 0.1% BSA (bovine serum albumin) or HSA (human serum albumin) to the PBS to prevent protein adsorption and loss, especially at low concentrations.
• Gently add the buffer to the vial. Do not vortex; instead, gently swirl or pipette up and down to dissolve. Let the vial stand at room temperature for 15–30 minutes with gentle agitation until fully dissolved.
• If undissolved particulates remain, mix gently for up to 2 hours at room temperature, or overnight at 4 °C.

3. Dilution for Cell Culture

• After initial reconstitution, dilute the stock solution to the desired working concentration using cell culture medium or PBS containing carrier protein (e.g., 0.1% BSA or 10% FBS).
• Avoid diluting in plain buffer without carrier protein, as this can lead to significant protein loss due to adsorption to plasticware.

4. Aliquoting and Storage

• Aliquot the reconstituted protein into single-use volumes (≥20 μL) to avoid repeated freeze-thaw cycles, which reduce activity.
• For short-term storage (up to 1 week), keep at 2–8 °C under sterile conditions.
• For longer-term storage (up to 3 months), store at –20 °C to –70 °C under sterile conditions.
• For extended storage, consider adding 5–50% glycerol as a cryoprotectant before freezing.

5. General Best Practices

• Always consult the product’s Certificate of Analysis or datasheet for lot-specific instructions, as formulations may vary.
• Use sterile technique throughout to prevent contamination.
• Avoid vigorous agitation or vortexing, which can denature the protein.

Summary Table: Key Steps for Recombinant Human Lymphotactin Reconstitution

Typical working concentrations for cell culture are in the range of 0.08–0.5 μg/mL, but optimize based on your specific assay.

If your experiment is serum-free or animal-derived protein-free, use trehalose as a stabilizer instead of BSA or FBS.

These steps will ensure that your recombinant human lymphotactin is properly reconstituted and retains its biological activity for cell culture applications.