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RB545 Rat Anti-Mouse Ly-6G
RB545 Rat Anti-Mouse Ly-6G
Flow cytometric analysis of Ly-6G expression on Mouse bone marrow cells. C57BL/6 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™ RB545 Rat IgG2a, κ Isotype Control (Cat. No. 568130​; dashed line histogram) or BD Horizon™ RB545 Rat Anti-Mouse Ly-6G antibody (Cat. No. 569726/569754; solid line histogram) at 0.5 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing the expression of Ly-6G (or Ig Isotype control staining) was derived from gated events with the forward and side-light scatter characteristics of viable (DAPI-negative) bone marrow cells. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 software. Data shown on this Technical Data Sheet are not lot specific.
RB545 Rat Anti-Mouse Ly-6G
Multiparameter flow cytometric analysis of Ly-6G expression on Mouse bone marrow cells. C57BL/6 Mouse bone marrow cells were stained with either BD Horizon™ RB545 Rat IgG2a, κ Isotype Control (Cat. No. 568130; Left Plot) or BD Horizon™ RB545 Rat Anti-Mouse Ly-6G antibody (Cat. No. 569726/569754; Right Plot) at 0.5 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 Ly-6G (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) bone marrow leukocyte populations. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of Ly-6G expression on Mouse bone marrow cells. C57BL/6 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™ RB545 Rat IgG2a, κ Isotype Control (Cat. No. 568130​; dashed line histogram) or BD Horizon™ RB545 Rat Anti-Mouse Ly-6G antibody (Cat. No. 569726/569754; solid line histogram) at 0.5 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing the expression of Ly-6G (or Ig Isotype control staining) was derived from gated events with the forward and side-light scatter characteristics of viable (DAPI-negative) bone marrow cells. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of Ly-6G expression on Mouse bone marrow cells. C57BL/6 Mouse bone marrow cells were stained with either BD Horizon™ RB545 Rat IgG2a, κ Isotype Control (Cat. No. 568130; Left Plot) or BD Horizon™ RB545 Rat Anti-Mouse Ly-6G antibody (Cat. No. 569726/569754; Right Plot) at 0.5 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 Ly-6G (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) bone marrow leukocyte populations. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Horizon™
Ly6g; Lymphocyte antigen 6G; Lymphocyte antigen 6 complex, locus G; Gr1
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2a, κ
Ly-6G-transfected EL4J Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
546644
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

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 unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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.

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. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. For U.S. patents that may apply, see bd.com/patents.
569726 Rev. 2
Antibody Details
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1A8

The 1A8 monoclonal antibody specifically binds to Ly-6G, a 21-25-kDa GPI-anchored protein. In the bone marrow, Ly6G is expressed on the majority of the largest cells, predominantly granulocytes, but not on lymphoid or erythroid cells.  In the periphery, it is expressed on granulocytes. The mAb RB6-8C5 recognizes both Ly-6G and Ly-6C and blocks the binding of mAb 1A8 to Ly-6G.

569726 Rev. 2
Format Details
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RB545
The BD Horizon RealBlue™ 545 (RB545) fluorochrome is part of the BD family of blue (488-nm) laser dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 496-nm and an emission maximum (Em Max) at 545-nm. Driven by BD innovation, RB545 can be used on spectral cytometers and is designed to be excited by the blue (488-nm) laser with minimal excitation by the 561-nm yellow-green laser. RB545 has minimal spillover into yellow-green detectors and a brightness level similar to FITC.  Given its unique emission max, on a spectral instrument RB545 can be used simultaneously with FITC and PE to provide an additional color off of the blue laser. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
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RB545
Blue 488 nm
496 nm
545 nm
569726 Rev.2
Citations & References
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Development References (5)

  1. 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. (Immunogen: Flow cytometry, Immunoprecipitation). View Reference
  2. Fleming TJ, Malek TR. Multiple glycosylphosphatidylinositol-anchored Ly-6 molecules and transmembrane Ly-6E mediate inhibition of IL-2 production. J Immunol. 1994; 153(5):1955-1962. (Clone-specific: Flow cytometry, Functional assay, Inhibition). View Reference
  3. Fleming TJ, O'HUigin C, Malek TR. Characterization of two novel Ly-6 genes. Protein sequence and potential structural similarity to alpha-bungarotoxin and other neurotoxins. J Immunol. 1993; 150(12):5379-5390. (Biology). View Reference
  4. Han G, Havnaer A, Lee HH, Carmichael DJ, Martinez LR. Biological depletion of neutrophils attenuates pro-inflammatory markers and the development of the psoriatic phenotype in a murine model of psoriasis.. Clin Immunol. 2020; 210:108294. (Clone-specific: Blocking, Flow cytometry). View Reference
  5. Nowroozilarki N, Öz HH, Schroth C, et al. Anti-inflammatory role of CD11b+Ly6G+ neutrophilic cells in allergic airway inflammation in mice.. Immunol Lett. 2018; 204:67-74. (Clone-specific: Flow cytometry). View Reference
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569726 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.