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Flow cytometric analysis of CD97 v2 expression on mouse splenocytes. BALB/c mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. No. 553141 or 553142). The cells were then stained with either BD Horizon™ BV421 Rat IgG1, κ Isotype Control (Cat. No. 562868; dashed line histogram) or BD OptiBuild™ BV421 Rat Anti-Mouse CD97 v2 (Cat. No. 747935; solid line histogram) at 0.25 μg/test. The fluorescence histogram showing the expression of CD97 v2 (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. The above is qualification data only and does not represent a specific OptiBuild™ lot.


BD OptiBuild™ BV421 Rat Anti-Mouse CD97 v2

Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads 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).
Product Notices
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
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The 587702 monoclonal antibody specifically recognizes the alternately-spliced variant 2 of CD97 (CD97 v2). CD97 is an N-glycosylated seven-span transmembrane protein that is encoded by Adgre5 (Adhesion G protein-coupled receptor E5) which belongs to the EGF-TM7 subfamily within the G-protein coupled receptor (GPCR) II family. Several CD97 isoforms have been described that have either three or four epidermal growth factor-like (EGF) domains in their N-terminal extracellular region. Compared with the longest form, CD97 v2 has a 94-amino acid deletion that results in the loss of the third EGF-like repeat. CD97 is expressed on monocytes, macrophages, T cells, B cell subsets, dendritic cells, granulocytes, hematopoietic stem cells and progenitor cells, and smooth muscle cells. Through its EGF domains, CD97 binds to glycophosphatidylinositol-linked CD55 that is also known as decay accelerating factor (DAF) and is involved in the regulation of complement activity. CD97 plays a role in angiogenesis, neutrophil migration, and host defense.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.

Development References (3)
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Carver EA, Hamann J, Olsen AS, Stubbs L. Physical mapping of EMR1 and CD97 in human Chromosome 19 and assignment of Cd97 to mouse Chromosome 8 suggest an ancient genomic duplication.. Mamm Genome. 1999; 10(10):1039-40. (Biology). View Reference
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Kwakkenbos MJ, Kop EN, Stacey M, et al. The EGF-TM7 family: a postgenomic view.. Immunogenetics. 2004; 55(10):655-66. (Biology). View Reference
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Leemans JC, te Velde AA, Florquin S, et al. The epidermal growth factor-seven transmembrane (EGF-TM7) receptor CD97 is required for neutrophil migration and host defense. J Immunol. 2004, January; 172(2):1125-1131. (Biology). View Reference
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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.
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