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APC-R700 Rat Anti-Mouse Ly-6G Ly-6C
APC-R700 Rat Anti-Mouse Ly-6G Ly-6C
Two-parameter flow cytometric analysis of Ly-6G and Ly-6C expression on mouse bone-marrow cells. Mouse bone-marrow cells 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™ APC-R700 Rat IgG2b, κ Isotype Control (Cat. No. 564984, Left Plot) or BD Horizon APC-R700 Rat Anti-Mouse Ly-6G and Ly-6C antibody (Cat. No. 565510; Right Plot). Two-parameter flow cytometric contour plots showing the correlated expression of Ly-6G and Ly-6C expression (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System. For optimal flow cytometric analysis, we suggest that this reagent be titrated starting at less than or equal to 0.25 µg per million cells in a 100 µl volume.
Two-parameter flow cytometric analysis of Ly-6G and Ly-6C expression on mouse bone-marrow cells. Mouse bone-marrow cells 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™ APC-R700 Rat IgG2b, κ Isotype Control (Cat. No. 564984, Left Plot) or BD Horizon APC-R700 Rat Anti-Mouse Ly-6G and Ly-6C antibody (Cat. No. 565510; Right Plot). Two-parameter flow cytometric contour plots showing the correlated expression of Ly-6G and Ly-6C expression (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System. For optimal flow cytometric analysis, we suggest that this reagent be titrated starting at less than or equal to 0.25 µg per million cells in a 100 µl volume.
Product Details
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BD Horizon™
Ly6c, Lymphocyte antigen 6C2; Lymphocyte antigen 6G, Ly6g, Gr-1
Mouse (QC Testing)
Rat IgG2b, κ
Not Reported
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2739274
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 with BD Horizon APC-R700 under optimum conditions, and unconjugated antibody and free BD Horizon APC-R700 were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  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. An isotype control should be used at the same concentration as the antibody of interest.
565510 Rev. 1
Antibody Details
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RB6-8C5

The RB6-8C5 monoclonal antibody recognizes a common epitope on Ly-6G and Ly-6C, previously known as the myeloid differentiation antigen Gr-1. In the bone marrow, the level of antigen expression is directly correlated with granulocyte differentiation and maturation. The antigen is also expressed on the monocyte lineage in the bone marrow, but not on erythroid cells. In the periphery, RB6-8C5 antibody recognizes granulocytes (neutrophils and eosinophils) and monocytes. The RB6-8C5 antibody is a component of the "lineage cocktail" used in studies of hematopoietic cell lineages. The 1A8 antibody (Cat. No. 551461) specifically recognizes Ly-6G, but not Ly-6C.

Based on comparison of the staining patterns given by 1A8 versus RB6-8C5 antibodies on total blood leucocytes, it is evident that the 1A8 antibody stains the RB6-8C5-bright population, corresponding to Ly-6G-expressing granulocytes; whereas, the RB6-8C5-dim population is 1A8-negative and corresponds to Ly-6C-expressing lymphocytes and monocytes. Please refer to the Technical Data Sheets for Cat. No. 551459 and 553128 for more detailed information.

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).

565510 Rev. 1
Format Details
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APC-R700
The BD Horizon™ APC-R700 (APC-R700) Dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of an Allophycocyanin (APC) dye donor that has excitation maximum (Ex Max) of 651-nm and an acceptor dye, R700, with an emission maximum (Em Max) at 706-nm. APC-R700, driven by BD innovation, is designed to be excited by the red (627–640-nm) laser and detected using an optical filter centered near 710-nm (e.g., a 720/40-nm bandpass filter). APC-R700 is a brighter alternative to Alexa Fluor™ 700. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC-R700
Red 627-640 nm
651 nm
706 nm
565510 Rev.1
Citations & References
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Development References (11)

  1. Brummer E, Sugar AM, Stevens DA. Immunological activation of polymorphonuclear neutrophils for fungal killing: studies with murine cells and blastomyces dermatitidis in vitro. J Leukoc Biol. 1984; 36(4):505-520. (Clone-specific: Cytotoxicity). View Reference
  2. Conlan JW, North RJ. Neutrophils are essential for early anti-Listeria defense in the liver, but not in the spleen or peritoneal cavity, as revealed by a granulocyte-depleting monoclonal antibody. J Exp Med. 1994; 179(1):259-268. (Clone-specific: Depletion, Western blot). View Reference
  3. Czuprynski CJ, Brown JF, Maroushek N, Wagner RD, Steinberg H. Administration of anti-granulocyte mAb RB6-8C5 impairs the resistance of mice to Listeria monocytogenes infection. J Immunol. 1994; 152(4):1836-1846. (Clone-specific: Depletion, Western blot). View Reference
  4. Fleming TJ, Fleming ML, Malek TR. Selective expression of Ly-6G on myeloid lineage cells in mouse bone marrow. RB6-8C5 mAb to granulocyte-differentiation antigen (Gr-1) detects members of the Ly-6 family. J Immunol. 1993; 151(5):2399-2408. (Clone-specific: Immunoprecipitation). View Reference
  5. Gumley TP, McKenzie IF, Sandrin MS. Tissue expression, structure and function of the murine Ly-6 family of molecules. Immunol Cell Biol. 1995; 73(4):277-296. (Biology). View Reference
  6. Hestdal K, Ruscetti FW, Ihle JN, et al. Characterization and regulation of RB6-8C5 antigen expression on murine bone marrow cells. J Immunol. 1991; 147(1):22-28. (Biology). View Reference
  7. Lagasse E, Weissman IL. Flow cytometric identification of murine neutrophils and monocytes. J Immunol Methods. 1996; 197(1-2):139-150. (Biology). View Reference
  8. Lewinsohn DM, Bargatze RF, Butcher EC. Leukocyte-endothelial cell recognition: evidence of a common molecular mechanism shared by neutrophils, lymphocytes, and other leukocytes. J Immunol. 1987; 138(12):4313-4321. (Biology). View Reference
  9. Stoppacciaro A, Melani C, Parenza M, et al. Regression of an established tumor genetically modified to release granulocyte colony-stimulating factor requires granulocyte-T cell cooperation and T cell-produced interferon gamma. J Exp Med. 1993; 178(1):151-161. (Clone-specific: Depletion, Immunohistochemistry). View Reference
  10. Tepper RI, Coffman RL, Leder P. An eosinophil-dependent mechanism for the antitumor effect of interleukin-4. Science. 1992; 257(5069):548-551. (Biology). View Reference
  11. Tumpey TM, Chen SH, Oakes JE, Lausch RN. Neutrophil-mediated suppression of virus replication after herpes simplex virus type 1 infection of the murine cornea. J Virol. 1996; 70(2):898-904. (Clone-specific: Depletion). View Reference
View All (11) View Less
565510 Rev. 1

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