CCL26/Eotaxin-3 Antibody [Unconjugated]

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.

CCL26/Eotaxin‑3 was detected in immersion fixed paraffin-embedded sections of human colon (vasculature) using Goat Anti-Human CCL26/Eotaxin‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF653) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface on endothelial cells. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

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 4‑24 µg/mL.

• Reactivity: Hu
• Applications: WB, IHC, B/N
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

CCL26/Eotaxin-3 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human CCL26/Eotaxin‑3
  Thr24-Leu94
  Accession # Q9Y258
• Specificity: Detects CCL26/Eotaxin‑3 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CCL26
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Neutralization 10-25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CCL26/Eotaxin-3 Antibody [Unconjugated]

• CC chemokine IMAC
• C-C motif chemokine 26
• CCL26
• chemokine (C-C motif) ligand 26
• chemokine N1
• Eotaxin3
• Eotaxin-3
• IMAC
• Macrophage inflammatory protein 4-alpha
• MGC126714
• MIP-4a
• MIP-4alpha
• SCYA26
• SCYA26MIP-4-alpha
• small inducible cytokine A26
• small inducible cytokine subfamily A (Cys-Cys), member 26
• Small-inducible cytokine A26
• TSC-1Thymic stroma chemokine-1

Background

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.

1. Gou, R.F. et al. (1999) Genomics 58:313.
2. Kitamura, M. et al. (1999) J. Biol. Chem. 274:27975.
3. Shinkai, A. et al. (1999) J. Immunol. 163:1602.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CCL26/Eotaxin-3 Antibody (AF653)(5)

Publications using AF653

• Nagasaki T, Schuyler AJ, Zhao J Et al. 15LO1 dictates glutathione redox changes in asthmatic airway epithelium to worsen type-2 inflammation The Journal of clinical investigation 11/11/2021 [PMID: 34762602]
• Rump L, Mattey D, Kehoe O, Middleton J An initial investigation into endothelial CC chemokine expression in the human rheumatoid synovium. Cytokine, 2017-09-01;97(0):133-140. 2017-09-01 [PMID: 28648867] (IHC-Fr, Human)
• Da Hyun Jung, Gak-Won Yun, Yoo Jin Lee, Yunju Jo, Hyojin Park Clinicopathologic Analysis of Proton Pump Inhibitor-Responsive Esophageal Eosinophilia in Korean Patients Gut and Liver 2016-01-23 [PMID: 25963082]
• Park , Jason Y, Zhang , Xi, Nguyen , Nathalie, Souza , Rhonda F, Spechler , Stuart J, Cheng , Edaire Proton pump inhibitors decrease eotaxin-3 expression in the proximal esophagus of children with esophageal eosinophilia. PLoS ONE, 2014-07-02;9(7):e101391. 2014-07-02 [PMID: 24988451] (IHC, Human)
• Kagami S, Kai H, Kakinuma T High Levels of CCL26 in Blister Fluid and Sera of Patients with Bullous Pemphigoid. J. Invest. Dermatol., 2011-09-01;132(1):249-51. 2011-09-01 [PMID: 21881593] (IHC-P, Human)

Bioinformatics

• Gene Symbol: CCL26
• Uniprot:
  • Q9Y258()