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Alexa Fluor® 700 Rat Anti-Mouse CD24
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Alexa Fluor® 700 Rat Anti-Mouse CD24
Two-color flow cytometric analysis of CD24 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 Alexa Fluor® 700 Rat IgG2b, κ Isotype Control (Cat. No. 557964, Left Panel) or Alexa Fluor® 700 Rat Anti-Mouse CD24 antibody (Cat. No. 564237, Right Panel). Two-color flow cytometric dot plots showing the correlated expression patterns of CD24 (or Ig Isotype control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Two-color flow cytometric analysis of CD24 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 Alexa Fluor® 700 Rat IgG2b, κ Isotype Control (Cat. No. 557964, Left Panel) or Alexa Fluor® 700 Rat Anti-Mouse CD24 antibody (Cat. No. 564237, Right Panel). Two-color flow cytometric dot plots showing the correlated expression patterns of CD24 (or Ig Isotype control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
CD24a; HSA; Heat Stable Antigen; Ly-52; Nectadrin; R13-Ag
Mouse (QC Testing)
Rat DA, also known as DA/HA IgG2b, κ
C57BL/10 Mouse Splenic T Lymphocytes
Flow cytometry (Routinely Tested)
0.2 mg/ml
12484
AB_2738691
Aqueous buffered solution containing protein stabilizer and ≤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 Alexa Fluor® 700 under optimum conditions, and unreacted Alexa Fluor® 700 was removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  5. Alexa Fluor® 700 has an adsorption maximum of ~700nm and a peak fluorescence emission of ~720nm. Before staining cells with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564237 Rev. 1
Antibody Details
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M1/69

The M1/69 monoclonal antibody specifically binds to CD24 (Heat-Stable Antigen, HSA or HsAg), a variably glycosylated, glycosyl-phosphatidylinositol-anchored membrane protein expressed on erythrocytes, granulocytes, monocytes, lymphocytes, and neurons. Hematopoietic stem cells of the embryonic yolk sac and fetal liver express CD24. Levels of expression of CD24 vary during differentiation of the T and B cell lineages. In  the bone marrow, hematopoietic progenitors acquire CD24 expression upon commitment to the B-lymphocyte lineage. Immature B cells in the bone marrow express low CD24 levels whereas peripheral B lymphocytes express intermediate to high levels of CD24. The level of CD24 expression has been reported to rise upon activation of splenic B cells with LPS, but not with CD154 (CD40 Ligand). The majority of thymocytes express high levels of CD24, while most mature thymic and peripheral T lymphocytes do not express CD24. In contrast, TCR-bearing thymocytes which emigrate to the spleen are CD24+. Dendritic cells of the thymus, spleen, liver, and epidermal Langerhans cells have also been reported to express CD24. CD24 is not expressed by NK cells, as determined by staining with J11d mAb (Cat. No. 553146). CD24 is involved in the costimulation of CD4+ T cells by B cells, it is a "co-inducer" of in vitro thymocyte maturation, and it is a ligand of CD62P (P-selectin). While the monoclonal antibodies 30-F1, M1/69, and J11d all react with CD24, they show subtle differences in the level of staining of different lymphocyte populations. When possible, investigators should continue to use the same monoclonal anti-CD24 antibody as used in previous studies.

564237 Rev. 1
Format Details
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Alexa Fluor™ 700
Alexa Fluor™ 700 dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 697 nm and an emission maximum (Em Max) at 719-nm. Alexa Fluor™ 700 is designed to be excited by the Red (627–640-nm) laser and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 700
Red 627-640 nm
697 nm
719 nm
564237 Rev.1
Citations & References
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Development References (11)

  1. 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. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  2. Allman DM, Ferguson SE, Lentz VM, Cancro MP. Peripheral B cell maturation. II. Heat-stable antigen(hi) splenic B cells are an immature developmental intermediate in the production of long-lived marrow-derived B cells. J Immunol. 1993; 151(9):4431-4444. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  3. Alterman LA, Crispe IN, Kinnon C. Characterization of the murine heat-stable antigen: an hematolymphoid differentiation antigen defined by the J11d, M1/69 and B2A2 antibodies. Eur J Immunol. 1990; 20(7):1597-1602. (Clone-specific: Flow cytometry). View Reference
  4. 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. (Clone-specific: (Co)-stimulation, Flow cytometry). View Reference
  5. Crispe IN, Bevan MJ. Expression and functional significance of the J11d marker on mouse thymocytes. J Immunol. 1987 April; 138(7):2013-2018. (Clone-specific: Flow cytometry). View Reference
  6. Milstein C, Galfre G, Secher DS, Springer T. Monoclonal antibodies and cell surface antigens. Ciba Found Symp. 1979; 66:251-276. (Clone-specific: Flow cytometry). View Reference
  7. Springer T, Galfre G, Secher D, Milstein C. Monoclonal xenogeneic antibodies to mouse leukocyte antigens: identification of macrophage-specific and other differentiation antigens. Curr Top Microbiol Immunol. 1978; 81:45-50. (Immunogen: Radioimmunoassay). View Reference
  8. Springer T, Galfre G, Secher DS, Milstein C. Monoclonal xenogeneic antibodies to murine cell surface antigens: identification of novel leukocyte differentiation antigens. Eur J Immunol. 1978; 8(8):539-551. (Immunogen: Blocking, Radioimmunoassay). View Reference
  9. 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.
  10. Veillette A, Zuniga-Pflucker JC, Bolen JB, Kruisbeek AM. Engagement of CD4 and CD8 expressed on immature thymocytes induces activation of intracellular tyrosine phosphorylation pathways. J Exp Med. 1989; 170(5):1671-1680. (Clone-specific: Cell separation, Cytotoxicity). View Reference
  11. Wenger RH, Rochelle JM, Seldin MF, Kohler G, Nielsen PJ. The heat stable antigen (mouse CD24) gene is differentially regulated but has a housekeeping promoter. J Biol Chem. 1993; 268(31):23345-23352. (Biology). View Reference
View All (11) View Less
564237 Rev. 1

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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.