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Two color flow cytometric analysis of CD19 expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Hamster Anti-Mouse CD3e antibody (Cat. No. 553064/553063/561824) and either BD Horizon™ APC-R700 Rat IgG2a, κ Isotype Control (Cat. No. 564982; Left Plot) or BD Horizon APC-R700 Rat Anti-Mouse CD19 antibody (Cat. No. 565473; Right Plot). The two-color flow cytometric contour plot showing the correlated expression of CD19 (or Ig Isotype control staining) versus CD3e was derived from gated events with the forward and side light-scatter characteristic of viable splenic leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ APC-R700 Rat Anti-Mouse CD19
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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.
Preparation And Storage
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
The 1D3 antibody reacts with CD19, a B lymphocyte-lineage differentiation antigen. CD19, a 95-kDa transmembrance glycoprotein, is a member of the immunoglobulin superfamily and is expressed throughout B-lymphocyte development from the pro-B cell through the mature B-cell stages. Terminally differentiated plasma cells do not express CD19. On the surface of mature B cells, the CD19 molecule associates with CD21 (CR-2) and CD81 (TAPA-1), and this multimolecular complex synergizes with surface immunoglobulin to promote cellular activation. Studies with CD19-deficient mice have suggested that the level of CD19 expression affects the generation and maturation of B cells in the bone marrow and periphery. B-1 lineage B cells, also known as CD5+ B cells, are drastically reduced or absent in CD19-deficient mice. Increased levels of CD19 expression correlate with increased frequencies of peritonal and splenic B-1 cells and reduced numbers of conventional B lymphocytes in the periphery. CD19 participates in B-lymphocyte development, B-cell activation, maturation of memory B cells and regulation of tolerance. CD19 has also been detected on peritoneal mast cells, co-localized with CD21/CD35, and it is proposed to play a role in complement-mediated mast-cell activation.
This antibody was conjugated to BD Horizon APC-R700, which has been developed exclusively by BD Biosciences as a better alternative to Alexa Fluor® 700. APC-R700 excites and emits at similar wavelengths to Alexa Fluor® 700 yet exhibits significantly improved brightness. This dye can be excited by the red laser and detected with the same filter set as Alexa Fluor® (eg, 730/45-nm filter).
Development References (12)
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Engel P, Zhou LJ, Ord DC, Sato S, Koller B, Tedder TF. Abnormal B lymphocyte development, activation, and differentiation in mice that lack or overexpress the CD19 signal transduction molecule. Immunity. 1995; 3(1):39-50. (Biology). View Reference
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Fearon DT. The CD19-CR2-TAPA-1 complex, CD45 and signaling by the antigen receptor of B lymphocytes. Curr Opin Immunol. 1993; 5(3):341-348. (Biology). View Reference
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Gommerman JL, Oh DY, Zhou X, et al. A role for CD21/CD35 and CD19 in responses to acute septic peritonitis: a potential mechanism for mast cell activation. J Immunol. 2000; 165(12):6915-6921. (Biology). View Reference
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Inaoki M, Sato S, Weintraub BC, Goodnow CC, Tedder TF. CD19-regulated signaling thresholds control peripheral tolerance and autoantibody production in B lymphocytes. J Exp Med. 1997; 186(11):1923-1931. (Biology). View Reference
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Krop I, Shaffer AL, Fearon DT, Schlissel MS. The signaling activity of murine CD19 is regulated during cell development. J Immunol. 1996; 157(1):48-56. (Clone-specific: Activation, Calcium Flux, (Co)-stimulation, Flow cytometry, Functional assay, Immunoprecipitation). View Reference
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Krop I, de Fougerolles AR, Hardy RR, Allison M, Schlissel MS, Fearon DT. Self-renewal of B-1 lymphocytes is dependent on CD19. Eur J Immunol. 1996; 26(1):238-242. (Immunogen: Flow cytometry, Fluorescence activated cell sorting, Functional assay, Immunoprecipitation, In vivo exacerbation). View Reference
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Rickert RC, Rajewsky K, Roes J. Impairment of T-cell-dependent B-cell responses and B-1 cell development in CD19-deficient mice. Nature. 1995; 376(6538):352-355. (Biology). View Reference
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Sato S, Jansen PJ, Tedder TF. CD19 and CD22 expression reciprocally regulates tyrosine phosphorylation of Vav protein during B lymphocyte signaling. Proc Natl Acad Sci U S A. 1997; 94(24):13158-13162. (Biology). View Reference
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Sato S, Miller AS, Howard MC, Tedder TF. Regulation of B lymphocyte development and activation by the CD19/CD21/CD81/Leu 13 complex requires the cytoplasmic domain of CD19. J Immunol. 1997; 159(7):3278-3287. (Biology). View Reference
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Sato S, Ono N, Steeber DA, Pisetsky DS, Tedder TF. CD19 regulates B lymphocyte signaling thresholds critical for the development of B-1 lineage cells and autoimmunity. J Immunol. 1996; 157(10):4371-4378. (Biology). View Reference
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Sato S, Steeber DA,Jansen PJ, Tedder TF. CD19 expression levels regulate B lymphocyte development: human CD19 restores normal function in mice lacking endogenous CD19. J Immunol. 1997; 158(10):4662-4669. (Biology). View Reference
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Tedder TF, Zhou LJ, Engel P. The CD19/CD21 signal transduction complex of B lymphocytes. Immunol Today. 1994; 15(9):437-442. (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.