Detection of Human CCL26/Eotaxin‑3 by Western Blot. Western blot shows lysates of HUVEC human umbilical vein endothelial cells untreated (-) or treated (+) with 100 U/mL Recombinant Human IL‑4 (Catalog # 204-IL) for 24 hours. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human CCL26/Eotaxin‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF653) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CCL26/|
Eotaxin‑3 at approximately 12 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
|Chemotaxis Induced by CCL26/Eotaxin‑3 and Neutralizatio by Human CCL26/Eotaxin‑3 Antibody. Recombinant Human CCL26/Eotaxin‑3 (Catalog # 346‑E3) chemoattracts the BaF3 mouse pro‑B cell line transfected with mouse CCR3 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 CCL26/Eotaxin‑3 (1 µg/mL) is neutralized (green line) by increasing concentrations of Human CCL26/Eotaxin‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF653). The ND50 is typically 10‑25 µg/mL.|
Eotaxin-3, also named CCL26 or SCYA26, is a novel human CC chemokine that maps to chromosome 7q11.2, within 40 kilobases of the Eotaxin-2 loci. Eotaxin‑3/CCL26 has been shown to be constitutively expressed in the heart and ovary. In addition, low levels of Eotaxin-3/CCL26 expression can also be detected in various tissues. The expression of Eotaxin-3/CCL26 in vascular endothelial cells has been shown to be up-regulated by IL-13 and IL-4.
Eotaxin-3/CCL26 cDNA encodes a 94 amino acid (aa) residue protein with a putative signal peptide of either 23 or 26 aa residues. Recombinant Eotaxin-3/CCL26 has been produced in insect cells using a baculovirus expression system and shown to contain 71 aa residues. Recombinant Eotaxin-3/CCL26 is chemotactic for eosinophils and PHA-activated T cells. Eotaxin-3/CCL26 induces calcium flux in eosinophils as well as in CCR3-transfected cells. Eotaxin-3/CCL26 has also been shown to cross-desensitize cells to other CCR3 ligands. Both the 71 aa residue and 68 aa residue variants of recombinant Eotaxin-3 have been expressed in E. coli and found to have equal potency in inducing chemotaxis of a human CCR3-transfected cell line.
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