Human CCL21/6Ckine Antibody

(11 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human CCL21/6Ckine in direct ELISAs and Western blots. In direct ELISAs, less than 15% cross-reactivity with recombinant mouse (rm) CCL21/6Ckine is observed and less than 5% cross-reactivity with recombinant human (rh) Fractalkine, rhTeck, rhMCP‑3, rmCRG‑2, rhVIC, and rmVIC is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human CCL21/6Ckine
    Ser24-Pro134
    Accession # Q6ICR7
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Human CCL21/6Ckine (Catalog # 366-6C)
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize CCL21/6Ckine-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CCR7. The Neutralization Dose (ND50) is typically 1-4 µg/mL in the presence of 50 ng/mL Recombinant Human CCL21/6Ckine.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Chemotaxis Induced by CCL21/6Ckine and Neutralization by Human CCL21/6Ckine Antibody. Recombinant Human CCL21/6Ckine (Catalog # 366‑6C) chemoattracts the BaF3 mouse pro‑B cell line transfected with human CCR7 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Human CCL21/6Ckine (50 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CCL21/6Ckine Antigen Affinity-purified Polyclonal Antibody (Catalog # AF366). The ND50 is typically 1‑4 µg/mL.
Immunocytochemistry
CCL21/6Ckine in Human PBMCs. CCL21/6Ckine was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using 10 µg/mL Goat Anti-Human CCL21/6Ckine Antigen Affinity-purified Polyclonal Antibody (Catalog # AF366) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: CCL21/6Ckine

6Ckine is a novel CC chemokine discovered independently by three groups from the EST database. 6Ckine, also named SLC (Secondary Lymphoid-tissue Chemokine), CCL21, and Exodus-2, shows 21-33% identity to other CC chemokines. 6Ckine contains the four conserved cysteines characteristic of beta chemokines plus two additional cysteines in its unusually long carboxyl-terminal domain. Human 6Ckine cDNA encodes a 134 amino acid (aa) residue, highly basic, precursor protein with a 23 aa residue signal peptide that is cleaved to form the predicted 111 aa residue mature protein. Mouse 6Ckine cDNA encodes a 133 aa residue protein with a 23 residue signal peptide that is cleaved to generate the 110 residue mature protein. Human and mouse 6Ckine are highly conserved, exhibiting 86% aa sequence identity. 6Ckine is constitutively expressed at high levels in lymphoid tissues such as lymph nodes, spleen, and appendix. In mouse, high levels of 6Ckine mRNA are also detected in the lung. The gene for human 6Ckine has been localized at human chromosome 9p13 rather than chromosome 17 where the genes of many human CC chemokines are clustered. The 6Ckine gene location is within a region of about 100 kb from the gene for MIP-3 beta /ELC, another novel CC chemokine.

Unlike most CC chemokines, 6Ckine is not chemotactic for monocytes. Recombinant mouse 6Ckine is chemotactic in vitro for thymocytes and activated T cells. Recombinant human 6Ckine has been shown to be chemotactic for some human T cell lines, resting PBL, and cultured T cells expanded with PHA and IL-2. 6Ckine has also been reported to inhibit hemopoietic progenitor colony formation in a dose-dependent manner. 6Ckine acts via a class of as yet unidentified CC receptors on both T cells and B cells that are not shared by any other CC chemokines so far tested.

  • References:
    1. Hedrick, J.A. and A. Zlotnik (1997) J. Immunol. 159:1589.
    2. Hromas, R. et al. (1997) J. Immunol. 159:2554.
    3. Nagira, M. et al. (1997) J. Biol. Chem. 272:19518.
  • Entrez Gene IDs:
    6366 (Human); 18829 (Mouse); 298006 (Rat)
  • Alternate Names:
    6Ckine; 6CkineSmall-inducible cytokine A21; Beta-chemokine exodus-2; CCL21; chemokine (C-C motif) ligand 21; CKb9; exodus-2; member 21; SCYA21; SCYA21MGC34555; secondary lymphoid tissue chemokine; SLC; SLCSecondary lymphoid-tissue chemokine; TCA-4
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

11 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Intracellular coexpression of CXC- and CC- chemokine receptors and their ligands in human melanoma cell lines and dynamic variations after xenotransplantation.
    Authors: Pinto S, Martinez-Romero A, O'Connor J, Gil-Benso R, San-Miguel T, Terradez L, Monteagudo C, Callaghan R
    BMC Cancer, 2014;14(0):118.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC Fresh
  2. Feasibility of the use of combinatorial chemokine arrays to study blood and CSF in multiple sclerosis.
    Authors: Edwards, Keith R, Goyal, Jaya, Plavina, Tatiana, Czerkowicz, Julie, Goelz, Susan, Ranger, Ann, Cadavid, Diego, Browning, Jeffrey
    PLoS ONE, 2013;8(11):e81007.
    Species: Human
    Sample Type: CSF
    Application: immunoassay
  3. Inflammation-induced secretion of CCL21 in lymphatic endothelium is a key regulator of integrin-mediated dendritic cell transmigration.
    Authors: Johnson LA, Jackson DG
    Int. Immunol., 2010;22(10):839-49.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. CCL21 overexpressed on lymphatic vessels drives thymic hyperplasia in myasthenia.
    Authors: Berrih-Aknin S, Ruhlmann N, Bismuth J, Cizeron-Clairac G, Zelman E, Shachar I, Dartevelle P, de Rosbo NK, Le Panse R
    Ann. Neurol., 2009;66(4):521-31.
    Species: Human
    Sample Type: Tissue Homogenates
    Application: ELISA Development
  5. CXCL12 is essential for migration of activated Langerhans cells from epidermis to dermis.
    Authors: Ouwehand K, Santegoets SJ, Bruynzeel DP, Scheper RJ, de Gruijl TD, Gibbs S
    Eur. J. Immunol., 2008;38(11):3050-9.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  6. Lymphatic precollectors contain a novel, specialized subpopulation of podoplanin low, CCL27-expressing lymphatic endothelial cells.
    Authors: Wick N, Haluza D, Gurnhofer E, Raab I, Kasimir MT, Prinz M, Steiner CW, Reinisch C, Howorka A, Giovanoli P, Buchsbaum S, Krieger S, Tschachler E, Petzelbauer P, Kerjaschki D
    Am. J. Pathol., 2008;173(4):1202-9.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Frozen
  7. Autologous chemotaxis as a mechanism of tumor cell homing to lymphatics via interstitial flow and autocrine CCR7 signaling.
    Authors: Shields JD, Fleury ME, Yong C, Tomei AA, Randolph GJ, Swartz MA
    Cancer Cell, 2007;11(6):526-38.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  8. Oncostatin M enhances CCL21 expression by microvascular endothelial cells and increases the efficiency of dendritic cell trafficking to lymph nodes.
    Authors: Sugaya M, Fang L, Cardones AR, Kakinuma T, Jaber SH, Blauvelt A, Hwang ST
    J. Immunol., 2006;177(11):7665-72.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  9. Cutting edge: nonproliferating mature immune cells form a novel type of organized lymphoid structure in idiopathic pulmonary fibrosis.
    Authors: Marchal-Somme J, Uzunhan Y, Marchand-Adam S, Valeyre D, Soumelis V, Crestani B, Soler P
    J. Immunol., 2006;176(10):5735-9.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Frozen
  10. Trafficking pathways and characterization of CD4 and CD8 cells recruited to the skin of humans experimentally infected with Haemophilus ducreyi.
    Authors: Humphreys TL, Baldridge LA, Billings SD, Campbell JJ, Spinola SM
    Infect. Immun., 2005;73(7):3896-902.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  11. Developing neonatal rabbit appendix, a primary lymphoid organ, is seeded by immature blood-borne B cells: evidence for roles for CD62L/PNAd, CCR7/CCL21, alpha4beta1 and LFA-1.
    Authors: Sinha RK, Mage RG
    Dev. Comp. Immunol., 2004;28(7):829-41.
    Species: Rabbit
    Sample Type: Whole Tissue
    Application: IHC
Expand to show all 11 Citations
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