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BB515 Rat Anti-Mouse CD24
BB515 Rat Anti-Mouse CD24

Multicolor flow cytometric analysis of CD24 expression on viable Mouse splenic leucocytes. C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with APC Rat Anti-Mouse CD19 antibody (Cat. No. 550992/561738) and with either BD Horizon™ BB515 Rat IgG2c, κ Isotype Control (Cat. No. 565091; Left Plot) or BD Horizon™ BB515 Rat Anti-Mouse CD24 antibody (Cat. No. 567811; Right Plot) at 1 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD24 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Multicolor flow cytometric analysis of CD24 expression on viable Mouse splenic leucocytes. C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with APC Rat Anti-Mouse CD19 antibody (Cat. No. 550992/561738) and with either BD Horizon™ BB515 Rat IgG2c, κ Isotype Control (Cat. No. 565091; Left Plot) or BD Horizon™ BB515 Rat Anti-Mouse CD24 antibody (Cat. No. 567811; Right Plot) at 1 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD24 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Horizon™
Cd24a; HSA; heat stable antigen; Ly-52; nectadrin
Mouse (QC Testing)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2c, κ
Mouse thymus or spleen
Flow cytometry (Routinely Tested)
0.2 mg/ml
12484
AB_3683923
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 and unconjugated antibody and free dye were removed.

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® CompBeads 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 BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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. 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.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Alexa Fluor™ is a trademark of Life Technologies Corporation.
567811 Rev. 1
Antibody Details
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30-F1

The 30-F1 monoclonal antibody specifically recognizes CD24 which is also known as Heat-Stable Antigen (HSA or HsAg). CD24 is a highly glycosylated sialoprotein that is glycosylphosphatidylinositol (GPI)-linked to the cell membrane. CD24 is encoded by Cd24a (CD24a antigen) and is variably expressed on thymocytes, lymphocytes, monocytes, granulocytes, and erythrocytes. Hematopoietic stem cells of the embryonic yolk sac and fetal liver express CD24. The expressed levels of CD24 vary during the developmental stages of cells within the T and B cell lineages. In the bone marrow, hematopoietic progenitors acquire CD24 expression upon commitment to the lymphocyte lineage. Immature B cells in the bone marrow and spleen of adult mice express high levels of CD24, whereas mature peripheral B cells express intermediate levels of CD24. The majority of thymocytes express high levels of CD24, whereas mature thymic and peripheral T cells do not express CD24. In contrast, γδ TCR-bearing thymocytes which emigrate to the spleen are CD24+. Dendritic cells of the thymus, spleen, and liver and epidermal Langerhans cells reportedly express CD24 whereas NK cells and plasma cells do not. CD24 can function as an adhesion molecule and serve as a ligand for CD62P (P-selectin). It can be involved in the costimulation of CD4+ T cells by B cells as well as function as a "co-inducer" of in vitro thymocyte maturation. 30-F1 and other CD24-specific monoclonal antibodies, such as, M1/69 and J11d, can show subtle differences in the staining patterns for different lymphocyte populations. For this reason, the consistent use of the same CD24-specific antibody is recommended during research studies.

567811 Rev. 1
Format Details
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BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BB515
Blue 488 nm
490 nm
515 nm
567811 Rev.1
Citations & References
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View product citations for antibody "567811" on CiteAb

Development References (15)

  1. Aigner S, Ruppert M, Hubbe M, et al. Heat stable antigen (mouse CD24) supports myeloid cell binding to endothelial and platelet P-selectin. Int Immunol. 1995; 7(10):1557-1565. (Biology). View Reference
  2. Allman DM, Ferguson SE, Cancro MP. Peripheral B cell maturation. I. Immature peripheral B cells in adults are heat-stable antigenhi and exhibit unique signaling characteristics. J Immunol. 1992; 149(8):2533-2540. (Biology). View Reference
  3. Ardavin C, Wu L, Ferrero I, Shortman K. Mouse thymic dendritic cell subpopulations. Immunol Lett. 1993; 38(1):19-25. (Biology). View Reference
  4. Auerbach R, Huang H, Lu L. Hematopoietic stem cells in the mouse embryonic yolk sac. Stem Cells. 1996; 14(3):269-280. (Biology). View Reference
  5. Bruce J, Symington FW, McKearn TJ, Sprent J. A monoclonal antibody discriminating between subsets of T and B cells. J Immunol. 1981; 127(6):2496-2501. (Biology). View Reference
  6. Cibotti R, Punt JA, Dash KS, Sharrow SO, Singer A. Surface molecules that drive T cell development in vitro in the absence of thymic epithelium and in the absence of lineage-specific signals. Immunity. 1997; 6(3):245-255. (Biology). View Reference
  7. Hardy RR, Carmack CE, Shinton SA, Kemp JD, Hayakawa K. Resolution and characterization of pro-B and pre-pro-B cell stages in normal mouse bone marrow. J Exp Med. 1991; 173(5):1213-1225. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  8. Hunte BE, Capone M, Zlotnik A, Rennick D, Moore TA. Acquisition of CD24 expression by Lin-CD43+B220(low)ckit(hi) cells coincides with commitment to the B cell lineage. Eur J Immunol. 1998; 28(11):3850-3856. (Biology). View Reference
  9. Kelly KA, Pearse M, Lefrancois L, Scollay R. Emigration of selected subsets of gamma delta + T cells from the adult murine thymus. Int Immunol. 1993; 5(4):331-335. (Biology). View Reference
  10. Ledbetter JA, Herzenberg LA. Xenogeneic monoclonal antibodies to mouse lymphoid differentiation antigens. Immunol Rev. 1979; 47:63-90. (Biology). View Reference
  11. Li YS, Hayakawa K, Hardy RR. The regulated expression of B lineage associated genes during B cell differentiation in bone marrow and fetal liver. J Exp Med. 1993; 178(3):951-960. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  12. Reichlin A, Yokoyama WM. Natural killer cell proliferation induced by anti-NK1.1 and IL-2. Immunol Cell Biol. 1998; 76(2):143-152. (Biology). View Reference
  13. Stall AM, Wells SM. FACS analysis of murine B-cell populations. In: Herzenberg LA, Weir DM, Blackwell C, ed. Weir's Handbook of Experimental Immunology. Blackwell Science Publishers; 1997:63.1-63.17.
  14. Vremec D, Zorbas M, Scollay R, et al. The surface phenotype of dendritic cells purified from mouse thymus and spleen: investigation of the CD8 expression by a subpopulation of dendritic cells. J Exp Med. 1992; 176(1):47-58. (Biology). View Reference
  15. Wilson A, Day LM, Scollay R, Shortman K. Subpopulations of mature murine thymocytes: properties of CD4-CD8+ and CD4+CD8- thymocytes lacking the heat-stable antigen. Cell Immunol. 1988; 117(2):312-326. (Biology). View Reference
View All (15) View Less
567811 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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.