Recombinant Human CCL21 (Mucin)

Recombinant Human CCL21 (Mucin)

Product No.: C1365

[product_table name="All Top" skus="C1365"]

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Alternate Names
6Ckine, CKb9, ECL, MGC34555, SCYA21, SLC, TCA4, Exodus-2
Product Type
Recombinant Protein
Expression Host
NS0 Cells
Species
Human

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Background

Chemokine (C-C motif) ligand 21 (CCL21) is a small cytokine belonging to the CC chemokine family. This chemokine is also known as 6Ckine (because it has six conserved cysteine residues instead of the four cysteines typical to chemokines), exodus-2, and secondary lymphoid-tissue chemokine (SLC). CCL21 elicits its effects by binding to a cell surface chemokine receptor known as CCR7.

In the cancer biology CCL21 seems to have multifaceted roles. CCL21 attracts CCR7 bearing cells especially T and dendritic cells but also various cancer cells. Besides the antitumour role as leukocyte recruiting, CCL21 has been shown to facilitate dendritic cell functions and to exert an angiostatic effect.

Human and mouse 6Ckine are highly conserved, exhibiting 86% amino acid sequence identity.

Protein Details

Purity
>95% by SDS-PAGE and analyzed by silver stain.
Endotoxin Level
<1.0 EU/µg as determined by the LAL method
Fusion Protein Tag
Histidine Tag
Protein Accession No.
NP_002980 & P78423
Amino Acid Sequence
(Human 6Ckine) sdggaqdccl kysqrkipak vvrsyrkqep slgcsipail flprkrsqae lcadpkelwv qqlmqhldkt pspqkpaqgc rkdrgasktg kkgkgskgck rtersqtpkg pgs, (Human Fractalkine mucin-like stalk) fekqigevkp rttpaaggmd esvvlepeat gesssleptp ssqeaqralg tspelptgvt gssgtrlppt pkaqdggpvg telfrvppvs taatwqssap hqpgpslwae aktseapstq dpstqastas spapeenaps egqrvwgqgq sprpensler eemgpvpaht dafqdwgpgs mahvsvvpvs segtpsrepv asgswtpkae epihatmdpq rlgvlitpvp daqaatrrqk qnhhhhhh
N-terminal Sequence Analysis
Ser24
State of Matter
Lyophilized
Predicted Molecular Mass
The predicted molecular weight of Recombinant Human 6Ckine is Mr 38.3 kDa. However, the actual molecular weight as observed by migration on SDS-PAGE is Mr 95-105 kDa.
Predicted Molecular Mass
38.3
Storage and Stability
This lyophilized protein is stable for six to twelve months when stored desiccated at -20°C to -70°C. After aseptic reconstitution, this protein may be stored at 2°C to 8°C for one month or at -20°C to -70°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles. See Product Insert for exact lot specific storage instructions.
Country of Origin
USA
Shipping
Next Day Ambient
NCBI Gene Bank

Leinco Protein Advisor

Powered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments.

Recombinant Human CCL21 (Mucin) is a valuable tool for research applications due to its multifaceted biological functions and well-characterized properties as a chemokine with significant immunological relevance.

Immune Cell Recruitment and Activation

CCL21 functions as a potent chemoattractant for specific immune cell populations, making it essential for studies involving immune cell trafficking and activation. The protein selectively stimulates chemotaxis of T lymphocytes and dendritic cells through binding to the CCR7 receptor. This chemotactic capability is particularly valuable because it facilitates the co-localization of antigen-stimulated dendritic cells and naïve T cells, allowing for T cell activation and induction of immune responses. The protein demonstrates an ED50 of 0.5-1.5 µg/mL for chemoattraction of human peripheral blood lymphocytes, providing quantifiable bioactivity for experimental design.

Dendritic Cell Enhancement

Beyond recruitment, CCL21 enhances dendritic cell functionality in multiple ways. Research demonstrates that the protein increases apoptotic resistance and endocytic capabilities of dendritic cells, while also improving their survival and cytotoxic activity. These properties make it particularly useful for studies investigating dendritic cell biology and their role in immune responses.

Cancer Immunotherapy Applications

CCL21 has emerged as a significant candidate for cancer immunotherapy research. The protein exhibits antitumor activity through leukocyte recruitment and can inhibit immunosuppressive neutrophil polarization by suppressing NF-κB pathway activation. Studies show that CCL21 enhances the therapeutic efficacy of immune checkpoint inhibitors, suggesting potential for combination immunotherapy approaches. Additionally, the protein exerts angiostatic effects, which may contribute to tumor growth inhibition.

Technical Advantages

The mucin-stalk chimera variant offers practical advantages for research. The protein is typically supplied as a lyophilized preparation with high purity (>95% by SDS-PAGE) and low endotoxin levels (<1.0 EU/µg). It demonstrates stability for 6-12 months when stored desiccated at -20°C to -70°C, and can be reconstituted for short-term storage at 2-8°C or long-term storage at -20°C to -70°C. The mucin-stalk chimera formulation has been specifically validated for bioactivity assays.

Translational Research Potential

The protein's effectiveness can be further enhanced through nanoformulation approaches, which enable sustained release and improved delivery to internal tumor sites. This opens possibilities for developing advanced delivery systems and studying prolonged immunological effects in complex tissue environments.

Human and mouse CCL21 sequences exhibit 86% amino acid identity, facilitating translational research between in vitro human studies and in vivo murine models.

You can use recombinant human CCL21 (mucin) as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately determined.

Key considerations:

  • Recombinant proteins are commonly used as ELISA standards when purified native protein is unavailable or impractical to obtain. Many commercial CCL21 ELISA kits specify that their standards are recombinant human CCL21, often produced in E. coli or mammalian systems.
  • The mucin stalk variant of CCL21 is a naturally occurring isoform, and several ELISA kits and recombinant protein suppliers offer this form specifically for use as a standard. The critical requirement is that the recombinant protein matches the epitope recognized by the capture and detection antibodies in your ELISA.
  • Purity and quantification: The recombinant CCL21 should be highly purified (typically >95% by SDS-PAGE) and its concentration accurately measured, ideally by absorbance at 280 nm using a known extinction coefficient or by amino acid analysis.
  • Carrier protein: Some recombinant standards are supplied with carrier proteins (e.g., BSA) to improve stability, which is generally acceptable for ELISA calibration.
  • Epitope compatibility: Confirm that the recombinant CCL21 (mucin) sequence includes the region(s) recognized by your ELISA antibodies. Most commercial kits are validated for both natural and recombinant forms, but if your assay uses custom antibodies, check for compatibility.

Best practices:

  • Prepare a standard curve with serial dilutions of the recombinant CCL21 (mucin) in the same buffer as your samples.
  • Run the standard curve in parallel with your samples in every assay to ensure accurate quantification.
  • If using a custom or in-house ELISA, validate the recombinant standard by comparing its signal to that of a known reference or by spike-and-recovery experiments.

Summary:
Recombinant human CCL21 (mucin) is suitable as an ELISA standard if it is pure, accurately quantified, and recognized by your assay antibodies. This approach is standard practice in quantitative ELISA protocols for CCL21.

Recombinant Human CCL21 (Mucin) has been validated primarily for bioactivity assays, especially for its chemotactic effects on lymphocytes, and has been widely used in research on immune cell migration, tumor immunology, and dendritic cell function.

Key validated applications in published research include:

  • Bioactivity/Chemotaxis Assays: The recombinant CCL21 (mucin stalk chimera) has been validated for its ability to chemoattract human peripheral blood lymphocytes in vitro, with a reported ED50 of 0.5–1.5 µg/mL. It is specifically chemotactic for T lymphocytes and dendritic cells, but not for B cells, macrophages, or neutrophils.
  • Tumor Immunology and In Situ Vaccination: CCL21 has been used in preclinical and clinical studies to enhance immune cell infiltration into tumors, particularly in the context of dendritic cell-based in situ vaccination for cancer immunotherapy. For example, CCL21-expressing dendritic cells have been shown to sensitize tumors to immune checkpoint inhibitors and promote tumor-specific immune memory in murine models of non-small cell lung cancer (NSCLC). These studies highlight its role in modulating the tumor microenvironment and enhancing antitumor immune responses.
  • Immune Cell Migration and Lymphoid Organ Homing: CCL21 is critical for directing the migration of CCR7+ cells (such as T cells and dendritic cells) to lymphoid tissues, and has been used in studies investigating lymphocyte homing, lymphoid neogenesis, and peripheral tolerance.
  • Angiostasis and Dendritic Cell Function: CCL21 has been shown to facilitate dendritic cell function and exert angiostatic effects, making it relevant for studies on leukocyte recruitment and vascular biology in the context of cancer and inflammation.

Additional research applications include:

  • In vitro studies of immune cell signaling and activation via CCR7.
  • Studies on lymphatic metastasis and immune surveillance in cancer models.

Summary Table: Validated Applications

Application AreaDescription/Validation Evidence
Bioactivity/ChemotaxisChemoattracts human lymphocytes in vitro
Tumor ImmunologyEnhances immune infiltration, ISV in cancer
Immune Cell Migration/HomingDirects CCR7+ cell migration to lymphoid tissue
Dendritic Cell FunctionFacilitates DC function, antigen presentation
AngiostasisInhibits angiogenesis in tumor models

Note: Most published research uses recombinant CCL21 (including mucin stalk chimeras) for mechanistic studies in immunology, cancer biology, and cell migration. The protein is typically validated for chemotactic bioactivity, and its use in animal models and in vitro systems is well documented.

To reconstitute and prepare Recombinant Human CCL21 (Mucin) protein for cell culture experiments, follow these steps:

  • Reconstitution:

    • Use sterile PBS (phosphate-buffered saline) containing at least 0.1% human or bovine serum albumin (BSA) as the reconstitution buffer to minimize protein adsorption and stabilize the protein.
    • A typical reconstitution concentration is 100 μg/mL. Adjust the volume of buffer added to the lyophilized protein to achieve this concentration.
    • If your protocol or assay requires a different concentration, you may dilute further with the same buffer.
  • Procedure:

    • Allow the vial and buffer to equilibrate to room temperature before opening to prevent condensation.
    • Briefly centrifuge the vial to collect all lyophilized material at the bottom.
    • Add the calculated volume of sterile PBS + 0.1% BSA gently down the side of the vial.
    • Gently mix by pipetting up and down or by gentle swirling. Avoid vigorous vortexing to prevent protein denaturation.
    • Let the solution stand at room temperature for 15–30 minutes with occasional gentle agitation to ensure complete dissolution.
    • If any visible particulates remain, continue gentle mixing for up to 2 hours at room temperature.
  • Storage after reconstitution:

    • Store the reconstituted protein at 2–8 °C for up to one month.
    • For longer-term storage, aliquot and freeze at –20 °C to –70 °C. Avoid repeated freeze-thaw cycles to maintain protein integrity.
    • If using for cell culture, always use aseptic technique to prevent contamination.
  • General notes:

    • If your experiment is sensitive to animal-derived components, use recombinant or human serum albumin.
    • For chemotaxis or functional assays, confirm the final working concentration based on published protocols or preliminary titration.

Summary Table: Reconstitution and Storage

StepRecommendation
BufferSterile PBS + 0.1% BSA
Concentration100 μg/mL (typical)
MixingGentle pipetting/swirl, 15–30 min at RT
Short-term storage2–8 °C, up to 1 month
Long-term storage–20 °C to –70 °C, aliquoted, avoid freeze-thaw cycles

These guidelines are consistent with best practices for recombinant chemokine handling and are suitable for cell culture applications.

Certificate of Analysis

IMPORTANT Use lot specific datasheet for all technical information pertaining to this recombinant protein.
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Prod No.
Description
C1365
Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.