Alexa Fluor® 700 Rat anti-Mouse Ly-6G and Ly-6C

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BD Pharmingen™ Alexa Fluor® 700 Rat anti-Mouse Ly-6G and Ly-6C

Name: Alexa Fluor® 700 Rat anti-Mouse Ly-6G and Ly-6C
Brand: BD Pharmingen™
SKU: 557979

Clone RB6-8C5

(RUO)

Two-parameter analysis of the expression of Ly-6G and/or Ly-6C on bone-marrow myeloid cells. C57BL/6 bone-marrow leukocytes were stained with either Alexa Fluor® 700-conjugated Rat IgG2b, k isotype control mAb A95-1 (Cat. no. 557964, Panel A) or Alexa Fluor® 700-conjugated mAb RB6-8C5 (Panel B) in the presence of Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. no. 553141/553142). Please note that the population of cells having the lowest SSC (erythroid and lymphoid cells) shows little expression of Ly-6G/Ly-6C, while most of the leukocytes with moderate-to-high SSC (myeloid cells) are Ly-6G/Ly-6C-positive. Flow cytometry was performed on a BD™ LSR II flow cytometry system.

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Product Details

Brand: BD Pharmingen™

Alternative Name: Ly6c, Lymphocyte antigen 6C2; Lymphocyte antigen 6G, Ly6g, Gr-1

Reactivity: Mouse (QC Testing)

Isotype: Rat IgG2b, κ

Immunogen: Not Reported

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

RRID: AB_396971

Storage Buffer: Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

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

Product Notices

Since applications vary, each investigator should titrate the reagent to obtain optimal results.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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.
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.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.

557979 Rev. 2

Antibody Details

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.

557979 Rev. 2

Format Details

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.

Format: Alexa Fluor™ 700

Excitation Source: Red 627-640 nm

Excitation Max: 697 nm

Emission Max: 719 nm

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Citations & References

Development References (6)

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). View Reference
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). View Reference
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
Lagasse E, Weissman IL. Flow cytometric identification of murine neutrophils and monocytes. J Immunol Methods. 1996; 197(1-2):139-150. (Clone-specific). View Reference
Osawa M, Tokumoto Y, Nakauchi H. Hematopoietic stem cells. In: Herzenberg LA, Weir DM, Blackwell C, ed. Weir's Handbook of Experimental Immunology, 5th Edition. Cambridge: Blackwell Science; 1996:66.1-66.5.
Tepper RI, Coffman RL, Leder P. An eosinophil-dependent mechanism for the antitumor effect of interleukin-4. Science. 1992; 257(5069):548-551. (Clone-specific). View Reference

View All (6)

557979 Rev. 2

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.