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BD Horizon™ BB700 Rat Anti-Mouse Ly-6G
Clone 1A8 (RUO)
Flow cytometric analysis of Ly-6G expression on mouse bone-marrow leucocytes. 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 stained with either BD Horizon™ BB700 Rat IgG2a, κ Isotype Control (Cat. No. 566413; dashed line histogram) or BD Horizon BB700 Rat Anti-Mouse LY-6G antibody (Cat. No. 566435/566453; solid line histogram) at 0.5 μg/ml. Cells were then stained with BD Via-Probe™ Red Nucleic Acid Stain (Cat. No. 565803/565804) and analyzed. The fluorescence histograms showing Ly-6G fluorescence (or Ig Isotype control staining) were derived from gated events with the light scattering characteristics of viable lymphoid (Left Panel) or myeloid (Right Panel) cells that were Via Probe Red-negative. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of Ly-6G expression on mouse bone-marrow leucocytes. 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 stained with either BD Horizon™ BB700 Rat IgG2a, κ Isotype Control (Cat. No. 566413; dashed line histogram) or BD Horizon BB700 Rat Anti-Mouse LY-6G antibody (Cat. No. 566435/566453; solid line histogram) at 0.5 μg/ml. Cells were then stained with BD Via-Probe™ Red Nucleic Acid Stain (Cat. No. 565803/565804) and analyzed. The fluorescence histograms showing Ly-6G fluorescence (or Ig Isotype control staining) were derived from gated events with the light scattering characteristics of viable lymphoid (Left Panel) or myeloid (Right Panel) cells that were Via Probe Red-negative. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of Ly-6G expression on mouse bone-marrow leucocytes. 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 stained with either BD Horizon™ BB700 Rat IgG2a, κ Isotype Control (Cat. No. 566413; dashed line histogram) or BD Horizon BB700 Rat Anti-Mouse LY-6G antibody (Cat. No. 566435/566453; solid line histogram) at 0.5 μg/ml. Cells were then stained with BD Via-Probe™ Red Nucleic Acid Stain (Cat. No. 565803/565804) and analyzed. The fluorescence histograms showing Ly-6G fluorescence (or Ig Isotype control staining) were derived from gated events with the light scattering characteristics of viable lymphoid (Left Panel) or myeloid (Right Panel) cells that were Via Probe Red-negative. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of Ly-6G expression on mouse bone-marrow leucocytes. 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 stained with either BD Horizon™ BB700 Rat IgG2a, κ Isotype Control (Cat. No. 566413; dashed line histogram) or BD Horizon BB700 Rat Anti-Mouse LY-6G antibody (Cat. No. 566435/566453; solid line histogram) at 0.5 μg/ml. Cells were then stained with BD Via-Probe™ Red Nucleic Acid Stain (Cat. No. 565803/565804) and analyzed. The fluorescence histograms showing Ly-6G fluorescence (or Ig Isotype control staining) were derived from gated events with the light scattering characteristics of viable lymphoid (Left Panel) or myeloid (Right Panel) cells that were Via Probe Red-negative. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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
AB_2739730
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 with BD Horizon™ BB700 under optimum conditions, and unconjugated antibody and free BD Horizon BB700 were removed.
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794 or 566349).
When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.
For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent 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
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
- 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.
- BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Brilliant Stain Buffer RUO
Size
100 Tests
Cat No.
563794
Brilliant Stain Buffer RUO
Size
1000 Tests
Cat No.
566349
Red Nucleic Acid Stain RUO
Size
0.1 ML
Cat No.
565803
BB700 Rat IgG2a, κ Isotype Control RUO
Size
50 µg
Cat No.
566413
566435 Rev. 1
Antibody Details
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.
The antibody was conjugated to BD Horizon BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes. It is a polymer-based tandem dye developed exclusively by BD Biosciences. With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.
566435 Rev. 1
Format Details
BB700
The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BB700
476 nm
695 nm
566435 Rev.1
Citations & References
View product citations for antibody "566435" on CiteAb
Development References (3)
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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
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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
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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
566435 Rev. 1
Please refer to Support Documents for Quality Certificates
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.