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Anti-Mouse CD197 (CCR7) Biotin

Also known as: EBI-1, CCR-7, MIP-3 beta Receptor

RUO: For Research Use Only

Staining of C57BL/6 splenocytes with Anti-Mouse CD4 FITC (cat. 11-0041) and 0.5 µg of Rat IgG2a κ Isotype Control Biotin (cat. 13-4321) (left) or 0.5 µg of Anti-Mouse CD197 (CCR7) Biotin (right) followed by Streptavidin PE (cat. 12-4317). Staining with Anti-Mouse CD197 (CCR7) was carried out at 37°C for 0.5 hours and Fc receptor binding was blocked by staining cells with Anti-Mouse CD16/CD32 Purified (cat. 14-0161). Cells in the lymphocyte gate were used for analysis.

Contents: Anti-Mouse CD197 (CCR7) Biotin
Catalog Number: 13-1971
Concentration: 0.5 mg/ml
Formulation: aqueous buffer, 0.09% sodium azide, contains stabilizer if necessary
Storage Conditions: Store at 2-8°C. Do not freeze.
Clone: 4B12
Host/Isotype: Rat IgG2a, κ

Product Options
Cat. No.	Size	Price	Add Qty to Cart
13-1971-63	10 ug	To view relevant pricing, please select your country and resume browsing.
13-1971-80	25 ug
13-1971-82	100 ug
13-1971-85	500 ug

Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.

Research Related Products
Cat. No.	Format
11-4317	Streptavidin FITC
12-4317	Streptavidin PE
13-4321	Rat IgG2a K Isotype Control Biotin
14-0161	Anti-Mouse CD16/CD32 Purified (93)
14-1972	Mouse CCL19-Fc Fusion Recombinant Protein
17-4317	Streptavidin APC

Available Formats
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications	Regulatory Status
12-1971	Anti-Mouse CD197 (CCR7) PE	488	575	FC	RUO
13-1971	Anti-Mouse CD197 (CCR7) Biotin	N/A	N/A	FC	RUO
14-1971	Anti-Mouse CD197 (CCR7) Purified	N/A	N/A	FC IP	RUO
16-1971	Anti-Mouse CD197 (CCR7) Functional Grade* Purified	N/A	N/A	FC IP	RUO
17-1971	Anti-Mouse CD197 (CCR7) APC	633	660	FC	RUO
25-1971	Anti-Mouse CD197 (CCR7) PE-Cy7	488	760	FC	RUO
45-1971	Anti-Mouse CD197 (CCR7) PerCP-Cy5.5	488	690	FC	RUO
48-1971	Anti-Mouse CD197 (CCR7) eFluor® 450 (Pacific Blue® replacement)	405	450	FC	RUO
56-1971	Anti-Mouse CD197 (CCR7) Alexa Fluor® 700	633	723	FC	RUO
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µL to 5 µL. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.

Description

Please see our website for continuous updates regarding the use of this product.

The 4B12 monoclonal antibody reacts with mouse CCR7, also known as EBI-1 and CD197. For optimal visualization of CCR7 expression on different cell types it is necessary to use multi-color staining to discriminate different cell subsets (please see Technical Data Sheets for recommended concentration for individual formats). To address specificity, the staining profile of 4B12 has been compared to a polyclonal antibody generated against a CCR7 peptide (Bjorkdahl et al). This analysis confirms that the polyclonal antibody and 4B12 stain similar populations of cells. Furthermore, 4B12 stains mouse CCR7-GFP fusion protein-transfected RBL cells (see data in cat. 14-1971).

CCR7 is a chemokine receptor for the chemokines CCL19 (CKß11, ELC, MIP3ß, Scya19, Exodus-3) and CCL21 (CKß9, SLC, MIP2ß, 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 (TCM) 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. This 4B12 antibody has been reported for use in flow cytometric analysis.

Applications Tested

The 4B12 antibody has been tested by flow cytometric analysis of C57BL/6, Balb/C and SJL/J splenocytes and thymocytes.

Important: Staining with the 4B12 monoclonal antibody requires different conditions than typically used for surface-antigen staining. Please use the protocol below. Moreover, we have found that staining at 37°C, rather than 4°C, results in brighter 4B12 staining, as well as better resolution between positive and negative populations. Please see data for the PE 4B12 (cat. 12-1971) which demonstrates a comparison of staining at 4°C and 37°C. Staining with 4B12 at 37°C is not expected to interfere with co-staining other antigens, however this should be evaluated for individual experiments.

1. Prepare cell suspension as normal and block FcγIIIR/FcγIIR with 5 μg/million cells purified anti-mouse CD16/32 (cat. 14-0161) for 15 minutes on ice. If red blood cell lysis is carried out as part of cell preparation, ensure that fixatives are not present in the red blood cell lysis solution as this will eliminate 4B12 staining.

2. Without washing, add 1 μg/million cells 4B12 and incubate in a 37°C waterbath or at 4°C (please see notes above) for 0.5 hours.

3. Wash cells 1X with 3 mls of Flow Cytometry Staining Buffer (cat. 00-4222) and decant supernatant.

4. Analyze cells on flow cytometer or proceed with secondary staining on ice as normal.

Note: Co-staining mouse CCR7 with the 4B12 antibody and the CCR7 ligand CCL19-Fc (cat. 14-1972) may be difficult due to different binding conditions required for the antibody versus the ligand, and steric hindrance which may prevent co-staining of 4B12 and CCL19-Fc. Cross-blocking experiments have demonstrated that 4B12 binding is able to prevent the detectable binding of CCL19-Fc, however not the opposite. Furthermore, the correlation between 4B12 and CCL19-Fc staining may be difficult to predict due to the presence of unknown CCL19-Fc receptors in addition to CCR7.

References

Ritter, U, Wiede, F, Mielenz, D, Kiafard, Z, Zwirner, J, and Korner H. 2004. Analysis of the CCR7 expression on murine bone arrow-derived and spleen dendritic cells. Journal of Leukocyte Biology. 76: 472-476. (4B12, FC, Pubmed)

Bjorkdahl O, Barber KA, Brett SJ, Daly MG, Plumpton C, Elshourbagy NA, Tite JP, Thomsen LL. 2003. Characterization of CC-chemokine receptor 7 expression on murine T cells in lymphoid tissues. Immunology. 110(2):170-9.

Waldner H, Sobel RA, et al. 2006. The autoimmune diabetes locus Idd9 regulates development of type 1 diabetes by affecting the homing of islet-specific T cells. J Immunol. 176(9):5455-62. (4B12, FC, PubMed)

Ohl L, Mohaupt M, Czeloth N, Hintzen G, Kiafard Z, Zwirner J, Blankenstein T, Henning G, Forster R. 2004. CCR7 governs skin dendritic cell migration under inflammatory and steady-state conditions. Immunity. 21(2):279-88. (4B12, FC, Pubmed)

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Unless indicated otherwise, all products are For Research Use Only. Not for use in diagnostic or therapeutic procedures. Not for further distribution without written consent. Not all products available in all regions.