Product Information

Contents: Affinity Purified Mouse CCL19-Fc (ELC, MIP3b, CKb11) for mouse CCR7 detection
Catalog Number: 14-1972
Sizes: 10 ug
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, 0.09% NaN₃, 0.2% BSA
Storage Conditions: Store at 4°C.
Avoid repeated freeze/thaw cycles. This product is guaranteed for 3 months upon receipt, when handled and stored as instructed.

BALB/C thymocytes (left) were stained with 0.06 μg of purified CCL19-Fc followed by PE anti-human Fcγ (cat. 12-4998), FITC anti-mouse CD4 (GK1.5) (cat. 11-0041) and APC anti-mouse CD8 (53-6.7) (cat. 17-0081). CCL19-Fc staining is shown in the histogram after gating on CD4+CD8+ (open histogram) and CD4+ (colored histogram) thymocytes.
BALB/C splenocytes (right) were stained with 0.06 μg of purified CCL19-Fc, followed by PE anti-human Fcγ and FITC anti-mouse CD4. Cells in the lymphocyte gate were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-1972	Purified Mouse CCL19-Fc (ELC, MIP3b, CKb11) for mouse CCR7 detection	N/A	N/A	FC

Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq.

Description

The purified mouse CCL19-Fc is a fusion protein of the mouse CCL19 chemokine with the human Fcγ fragment. The purified mouse CCL19-Fc can be used for the detection of mouse CCR7 expression when used with a fluorochrome-conjugated anti-human Fc antibody. CCR7 is a chemokine receptor for the chemokines CCL19 (CKβ11, ELC, MIP3β, Scya19, Exodus-3) and CCL21 (CKβ9, SLC, MIP2β, Scya21, Exodus-2). In recent years, the role of chemokines in directing the migration of lymphocytes has been well-characterized. One of the most important mediators of homeostatic trafficking of naïve T cells to secondary lymphoid organs (SLO) is the chemokine receptor CCR7. Binding of its ligands, CCL19 and CCL21, mediates the transendothelial migration of T cells across high endothelial venules into SLO. It has also been demonstrated that CCR7 plays a role in the localization of dendritic cells and B cells during an immune response.

In addition to its significant role in the chemotaxis of lymphocytes, human CCR7 has also been recognized as a marker for a distinct subset of memory T cells, the central memory (T_CM) population. These cells are characterized by the expression of CCR7 and CD62L and reside within peripheral lymphoid organs.

CCR7 also plays a role in thymocyte development and its deficiency leads to disturbed thymic architecture, aberrant T cell development, and limited thymocyte expansion.

Applications Reported

For research use only, not for diagnostic or therapeutic use. Purified Mouse CCL19-Fc has been reported for use in flow cytometric analysis.

Applications Tested

Recombinant purified mouse CCL19-Fc has been tested by flow cytometric analysis of mouse splenocytes and thymocytes. This can be used at less than or equal to 0.06 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

References

Hargreaves DC, Hyman PL, Lu TT, Ngo VN, Bidgol A, Suzuki G, Zou YR, Littman DR, Cyster JG. A coordinated change in chemokine responsiveness guides plasma cell movements. J Exp Med. 2001 Jul 2;194(1):45-56.

Misslitz A, Pabst O, Hintzen G, Ohl L, Kremmer E, Petrie HT, Forster R. Thymic T cell development and progenitor localization depend on CCR7. J Exp Med. 2004 Aug 16;200(4):481-91.

Junt T, Scandella E, Forster R, Krebs P, Krautwald S, Lipp M, Hengartner H, Ludewig B. Impact of CCR7 on priming and distribution of antiviral effector and memory CTL. J Immunol. 2004 Dec 1;173(11):6684-93.

Related Products

Cat. 12-1971    Phycoerythrin (PE) anti-mouse CCR7 (CD197, EBI-1) (clone 4B12)
Cat. 13-1971    Biotin anti-mouse CCR7 (CD197, EBI-1) (clone 4B12)
Cat. 14-1971    Affinity Purified anti-mouse CCR7 (CD197, EBI-1) (clone 4B12)
Cat. 17-1971    Allophycocyanin (APC) anti-mouse CCR7 (CD197, EBI-1) (clone 4B12)
Cat. 25-1971    Phycoerythrin-Cy7 (PE-Cy7) anti-mouse CCR7 (CD197, EBI-1) (clone 4B12)
Cat. 12-4998    Phycoerythrin (PE) Anti-Human IgG Fcg Fragment Specific, Minimal Reactivity to Mouse IgG (clone Polyclonal)
Cat. 13-4998    Biotin Anti-Human IgG Fcg Fragment Specific, Minimal Reactivity to Mouse IgG (clone Polyclonal)

Purified Mouse CCL19-Fc Staining Protocol:

Add 1 million cells/tube and block FcγIIIR/FcγIIR with 1 µg/million cells purified anti-mouse CD16/32 (Cat. No. 14-0161) for 15 minutes on ice.
Without washing, add 0.06 µg CCL19-Fc/million cells and incubate on ice for ½ hour. While cells are being stained with ELC-Fc, conjugated anti-human Fcγ (Cat. No. 12-4998) should be pre-adsorbed with 2% normal mouse serum (Cat. No. 24-5544) and 2% normal rat serum (Cat. No. 24-5555). Add normal mouse and rat serum to required volume of anti-human Fcγ to a final concentration of 2% and incubate on ice for ½ hour.
Wash cells with 3 ml Flow Staining Buffer, spin cells and decant supernatant.
Add pre-adsorbed, conjugated anti-human Fcγ to the cells and incubate on ice for 20 minutes.
Wash cells 1X with 3 ml of Flow Staining Buffer and decant supernatant.
Surface-stain cells as normal.
Analyze cells on flow cytometer.