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BUV661 Rat Anti-Mouse CD197 (CCR7)
Product Details
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BD OptiBuild™
C-C chemokine receptor type 7; CD197; Ccr7; Cmkbr7; EBI1; Ebi1h
Mouse (Tested in Development)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a
Mouse CCR7 Transfected Cell Line
Flow cytometry (Qualified)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Note:  When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed.  For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

Product Notices

  1. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. BD Horizon Brilliant Ultraviolet 661 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
741677 Rev. 1
Antibody Details
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4B12

The monoclonal antibody 4B12/CCR7 reacts with the mouse C-C chemokine receptor type 7 (CCR7). CCR7 is also known as CD197 (previously known as EBI1, Ebi1h and CMKBR7) and plays a central role in mediating homeostatic B and T lymphocyte trafficking to and within secondary lymphoid tissues. CD197 is a seven-transmembrane, G-protein-coupled, 43 kDa glycoprotein receptor that is specific for the CC chemokines, MIP3ß/Exodus-3/ELC/CKb11/Scya19/CCL19 and 6Ckine/Exodus-2/SLC/TCA4/CKb9/Scya21/CCL21. The mouse Ccr7 gene is located on chromosome 11. CD197 (CCR7) is differentially expressed by subsets of thymocytes. Positive CD197 expression appears to be involved in the cortex-to-medulla migration of positively-selected thymocytes wherein they complete functional maturation including the establishment of central tolerance. It is most highly expressed by some mature medullary single-positive thymocytes. CD197 is also expressed by subsets of mature peripheral CD4+ and CD8+ T lymphocytes including naïve and regulatory T cells and central memory T cells. In addition, it is differentially expressed by subsets of B lymphocytes, dendritic cells, and Langerhans cells. CD197 serves as a homing receptor that helps guide these various cell types to and within lymphoid tissues. In this way, CCR7 supports protective immunity while safeguarding self tolerance. Reportedly, the 4B12/CCR7 antibody is not agonistic, is not blocked by CCL21 nor by physiologic levels of CCL19, nor does the antibody block the binding of CCL21 to CCR7. The immunogen used to generate the 4B12 hybridoma was a mouse CCR7-transfected rat cell line.

The antibody was conjugated to BD Horizon BUV661 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 661-nm. BD Horizon Brilliant BUV661 can be excited by the ultraviolet laser (355 nm) and detected with a 670/25 filter and a 630 nm LP.  Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/25-nm filter).

741677 Rev. 1
Format Details
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BUV661
The BD Horizon Brilliant™ Ultraviolet 661 (BUV661) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 660-nm. BUV661, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 660-nm (e.g., 670/25 bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV661
Ultraviolet 355 nm
350 nm
660 nm
741677 Rev.1
Citations & References
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Development References (5)

  1. Britschgi MR, Link A, Lissandrin TK, Luther SA. Dynamic modulation of CCR7 expression and function on naive T lymphocytes in vivo. J Immunol. 2008; 181(11):7681-7688. (Clone-specific: Flow cytometry). View Reference
  2. Forster R, Davalos-Misslitz AC, Rot A. CCR7 and its ligands: balancing immunity and tolerance. Nat Rev Immunol. 2008; 8(5):362-371. (Biology). View Reference
  3. Kurobe, H., Liu, et al. CCR7-dependent cortex-to-medulla migration of positively selected thymocytes is essential for establishing central tolerance. Immunity. 2006; 24(2):165-177. (Biology). View Reference
  4. Ohl L, Mohaupt M, Czeloth N, et al. CCR7 governs skin dendritic cell migration under inflammatory and steady-state conditions. Immunity. 2004; 21(2):279-288. (Clone-specific: Flow cytometry, Immunofluorescence, Immunoprecipitation, Neutralization). View Reference
  5. Schweickart VL, Raport CJ, Godiska R, et al. Cloning of human and mouse EBI1, a lymphoid-specific G-protein-coupled receptor encoded on human chromosome 17q12-q21.2. Genomics. 1994; 23(3):643-650. (Biology). View Reference
View All (5) View Less
741677 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.