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Flow cytometric analysis of CD45.2 expression on C57BL/6 splenocytes. Splenic leucocytes from SJL mice (Left Panel) or C57BL/6 mice (Right Panel) were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either BD Horizon™ BV605 Mouse IgG2a, κ Isotype Control (Cat. No. 562778; dashed line histogram) or BD Horizon™ BV605 Mouse Anti-Mouse CD45.2 antibody (Cat. No. 563051; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.


BD Horizon™ BV605 Mouse Anti-Mouse CD45.2

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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
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Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
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The 104 monoclonal antibody recognizes the CD45 (Leukocyte Common Antigen) present on all leucocytes of most mouse strains (eg, A, AKR, BALB/c, CBA/Ca, CBA/J, C3H/He, C57BL, C57BR, C57L, C58, DBA/1, DBA/2, NZB, SWR, 129). This alloantigen was originally named Ly-5.1, and this was the designation at the time that the antibody was characterized. The designation was later changed from Ly-5.1 to Ly-5.2 to conform with the convention that the .2 alloantigen designations be assigned to the C57BL/6 strain. mAb 104 has been reported not to react with leucocytes of the mouse strains expressing the CD45.1 alloantigen (eg, RIII, SJL/J, STS/A, and DA). CD45 is a member of the Protein Tyrosine Phosphatase (PTP) family: its intracellular (COOH-terminal) region contains two PTP catalytic domains, and the extracellular region is highly variable due to alternative splicing of exons 4, 5, and 6 (designated A, B, and C, respectively), plus differing levels of glycosylation. The CD45 isoforms detected in the mouse are cell type-, maturation-, and activation state-specific. The CD45 isoforms play complex roles in T-cell and B-cell antigen receptor signal transduction. The 104 antibody has been reported to inhibit some responses of B cells, from mice expressing the CD45.2 alloantigen, to certain antigens and LPS. In addition, reduction of serum IgG levels and amelioration of autoimmune renal pathology were reported in mAb 104-treated systemic lupus erythematosus-prone mice.
This antibody is conjugated to BD Horizon BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.
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).

Development References (9)
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Johnson P, Greenbaum L, Bottomly K, Trowbridge IS. Identification of the alternatively spliced exons of murine CD45 (T200) required for reactivity with B220 and other T200-restricted antibodies. J Exp Med. 1989; 169(3):1179-1184. (Biology). View Reference
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Morse HC 3rd, Shen FW, Hammerling U. Genetic nomenclature for loci controlling mouse lymphocyte antigens. Immunogenetics. 1987; 25(2):71-78. (Biology). View Reference
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Ogimoto M, Mizuno K, Tate G, et al. Regulation of lipopolysaccharide- and IL-4-induced immunoglobulin heavy chain gene activation: differential roles for CD45 and Lyb-2. Int Immunol. 1992; 4(6):651-659. (Biology). View Reference
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Shen FW, Tung JS, Boyse EA. Further definition of the Ly-5 system. Immunogenetics. 1986; 24(3):146-149. (Biology). View Reference
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Shen FW. Monoclonal antibodies to mouse lymphocyte differentiation alloantigens. In: Hammerling GJ, Hammerling U, Kearney JF, ed. Monoclonal Antibodies and T-cell Hybridomas; Perspectives and Technical Advances. 1981:25-31.
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Yakura H, Ashida T, Kawabata I, Katagiri M. Alleviation of autoimmunity in BXSB mice by monoclonal alloantibody to Ly-5 (CD45). Eur J Immunol. 1989; 19(8):1505-1508. (Clone-specific: Inhibition, In vivo exacerbation). View Reference
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Yakura H, Kawabata I, Ashida T, Katagiri M. Differential regulation by Ly-5 and Lyb-2 of IgG production induced by lipopolysaccharide and B cell stimulatory factor-1 (IL-4). J Immunol. 1988; 141(3):875-880. (Clone-specific: Inhibition). View Reference
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Yakura H, Kawabata I, Shen FW, Katagiri M. Selective inhibition of lipopolysaccharide-induced polyclonal IgG response by monoclonal Ly-5 antibody. J Immunol. 1986; 136(8):2729-2733. (Clone-specific: Inhibition). View Reference
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Yakura H, Shen FW, Bourcet E, Boyse EA. On the function of Ly-5 in the regulation of antigen-driven B cell differentiation. Comparison and contrast with Lyb-2. J Exp Med. 1983; 157(4):1077-1088. (Clone-specific: Inhibition). View Reference
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