Recombinant Human CCL1

Recombinant Human CCL1 (also known as I-309) is a valuable tool for a wide range of research applications due to its well-characterized biological functions and its central role in immune regulation, inflammation, and disease pathogenesis. Here are several key reasons why you should consider using Recombinant Human CCL1 in your research:

1. Immune Cell Recruitment and Chemotaxis Studies

CCL1 is a potent chemoattractant for immune cells expressing its receptor, CCR8, including T cells, regulatory T cells (Tregs), and innate lymphoid cells (ILC2s). Using recombinant CCL1 allows you to:

• Study the migration and recruitment of CCR8+ cells in vitro and in vivo.
• Investigate the mechanisms of immune cell trafficking in inflammation, autoimmunity, and cancer.

2. Regulatory T Cell (Treg) Induction and Function

CCL1 has been shown to directly induce the differentiation of CD4+ Foxp3+ regulatory T cells, which are critical for immune tolerance and suppression of effector T cells. This makes recombinant CCL1 useful for:

• Studying Treg development and function.
• Exploring therapeutic strategies for autoimmune diseases, graft rejection, and graft-versus-host disease (GVHD).

3. Inflammation and Autoimmune Disease Research

CCL1 is upregulated in inflammatory conditions and plays a role in the pathogenesis of diseases such as HAM/TSP (HTLV-1-associated myelopathy/tropical spastic paraparesis), multiple sclerosis (MS), and atopic dermatitis. Recombinant CCL1 can be used to:

• Model inflammatory processes in vitro.
• Investigate the role of CCL1/CCR8 signaling in disease progression and potential therapeutic interventions.

4. Cancer Immunology and Tumor Microenvironment Studies

CCL1 is involved in the recruitment of immune cells to tumors and can influence the tumor microenvironment. Recombinant CCL1 is useful for:

• Studying the interaction between immune cells and tumor cells.
• Exploring strategies to enhance immune cell infiltration into tumors, such as in CAR T cell therapy for solid tumors.

5. Apoptosis, Angiogenesis, and Tumor Biology

CCL1 has been implicated in apoptosis, angiogenesis, and tumor biology. Recombinant CCL1 can be used to:

• Investigate the effects of CCL1 on cell survival and death.
• Study its role in tumor growth and metastasis.

6. High Purity and Low Endotoxin

Recombinant Human CCL1 is available with high purity (>97%) and low endotoxin levels (<1.0 EU/μg), ensuring reliable and reproducible results in sensitive assays and cell culture experiments.

7. Versatile Applications

Recombinant CCL1 can be used in a variety of applications, including:

• Bioassays for chemotaxis and cell migration.
• ELISA and other immunoassays.
• Cell culture and differentiation studies.
• In vivo animal models of disease.

8. Research Use Only

Recombinant Human CCL1 is intended for research use only, making it suitable for preclinical studies and experimental investigations without regulatory constraints.

In summary, Recombinant Human CCL1 is a versatile and well-characterized protein that can significantly enhance your research in immunology, inflammation, autoimmunity, cancer, and related fields. Its ability to modulate immune cell function and its involvement in various disease processes make it an essential tool for advancing our understanding of immune regulation and developing new therapeutic strategies.

Yes, recombinant human CCL1 is well-suited for use as a standard for quantification and calibration in ELISA assays. This is one of the primary recommended applications for recombinant CCL1 proteins.

Formulation Considerations

When selecting recombinant CCL1 for ELISA standardization, you have two main formulation options to consider:

Protein with Carrier (BSA-containing): This formulation includes bovine serum albumin as a carrier protein, which enhances protein stability, increases shelf-life, and allows storage at more dilute concentrations. This version is generally recommended for use as an ELISA standard.

Carrier-Free Formulation: This version lacks BSA and is recommended when the presence of carrier protein could interfere with your assay. However, carrier-free recombinant proteins are typically not recommended for bioassay applications, as they are not tested for such uses.

Typical Assay Ranges

Commercial ELISA kits using recombinant CCL1 standards operate across various detection ranges depending on the kit design: Assay ranges typically span from approximately 4-15.6 pg/mL at the lower detection limit to 1,000-3,000 pg/mL at the upper range, with sensitivities as low as 1.4-4 pg/mL depending on the specific kit configuration.

Reconstitution and Storage

Recombinant CCL1 is typically supplied in lyophilized form and should be reconstituted at 100 μg/mL in sterile phosphate-buffered saline (PBS) containing at least 0.1% human or bovine serum albumin. Store the reconstituted protein in a manual defrost freezer and avoid repeated freeze-thaw cycles to maintain stability and activity.

Recombinant Human CCL1 has been validated for several applications in published research, including bioassays (functional assays), ELISA (as a standard), blocking assays, Western blot, and cell culture experiments.

Key validated applications and supporting details:

• Bioassays / Functional Assays:
  Recombinant CCL1 is widely used to assess its chemoattractant activity on various immune cells, such as monocytes, NK cells, T cells, and dendritic cells, typically via migration or chemotaxis assays. For example, its ability to chemoattract BW5147 mouse T lymphoma cells or hCCR8-transfected BaF/3 cells is a standard functional validation.

• ELISA (Enzyme-Linked Immunosorbent Assay):
  CCL1 is used as a standard in ELISA to quantify CCL1 levels in biological samples, such as cell culture supernatants or bronchoalveolar lavage fluid (BALF). Published studies have measured CCL1 in human airway samples to investigate its role in asthma and other inflammatory conditions.

• Blocking Assays:
  Recombinant CCL1 is used in blocking assays to study the specificity of antibodies or to inhibit CCL1-CCR8 interactions in functional studies.

• Western Blot:
  The protein is validated for use as a positive control in Western blotting to detect CCL1 or to confirm antibody specificity.

• Cell Culture:
  Recombinant CCL1 is used to stimulate or recruit CCR8+ cells in vitro, including Treg migration assays and studies of immune cell differentiation or activation. For example, it has been used to demonstrate the recruitment of regulatory T cells (Tregs) in tumor models and to study T cell migration in cancer and inflammation.

• Research Areas:
  Applications span immunology, cancer research, inflammation, and autoimmune disease models. CCL1 has been used to study:

  • Treg recruitment in breast cancer and solid tumors
  • Immune cell migration in asthma and rheumatoid arthritis
  • Mechanisms of immune regulation and tumor microenvironment modulation

Summary Table of Validated Applications

In summary, recombinant human CCL1 is a versatile reagent validated for multiple immunological and biochemical applications, especially those involving chemotaxis, immune cell recruitment, and quantification in disease models.

To reconstitute and prepare Recombinant Human CCL1 protein for cell culture experiments, first check the product datasheet for specific instructions, as formulations may vary. The most common and recommended protocol is as follows:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Reconstitute the lyophilized protein at 100 μg/mL in sterile PBS (phosphate-buffered saline). If the datasheet specifies, add at least 0.1% carrier protein such as human or bovine serum albumin (BSA) to the PBS to stabilize the protein and prevent adsorption to plastic surfaces.
• Gently mix the solution by pipetting up and down or by gentle vortexing. Avoid foaming during reconstitution.
• Allow the protein to fully dissolve at room temperature for several minutes.
• If a different concentration is required, dilute the stock solution further in sterile PBS or cell culture medium, ideally containing 0.1–1% BSA or HSA as a carrier.
• Filter-sterilize the final working solution if sterility is required and the protein is not already sterile-filtered.

Storage after reconstitution:

• Store aliquots at 2–8 °C for up to 1 month or at –20 °C to –70 °C for longer-term storage.
• Avoid repeated freeze-thaw cycles to maintain protein integrity.

Additional notes:

• The typical working concentration for cell culture assays is in the ng/mL range (e.g., 1–10 ng/mL), but this should be optimized for your specific application.
• If the protein is supplied without carrier protein and you observe loss of activity or adsorption, consider adding BSA or HSA to your buffers.
• Always consult the specific product datasheet for any unique requirements, as some formulations may require reconstitution in water or other buffers.

Summary of key steps:

• Centrifuge vial → Reconstitute at 100 μg/mL in sterile PBS (+0.1% BSA if needed) → Mix gently → Aliquot and store appropriately → Dilute to working concentration in cell culture medium before use.

If your application is sensitive to endotoxin, confirm the endotoxin level is suitable for cell culture, especially for primary or sensitive immune cells.