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PerCP-Cy™5.5 Mouse Lineage Antibody Cocktail, with Isotype Control

PerCP-Cy™5.5 Mouse Lineage Antibody Cocktail, with Isotype Control

(RUO)
PerCP-Cy™5.5 Mouse Lineage Antibody Cocktail, with Isotype Control

Identification of CD34+ and CD34- subpopulations of hematopoietic progenitor cells. BALB/c bone marrow cells were treated with Mouse BD Fc Block™ Purified Rat anti-CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142) and stained with either FITC Rat IgG2a κ Isotype Control (Cat. No. 554688; data not shown) or FITC Rat anti-Mouse CD34 mAb RAM34 (Cat. No. 553733) and with either PerCP-Cy™ 5.5 Mouse Lineage Isotype Control Cocktail (Left Panel) or PerCP-Cy™ 5.5 Mouse Lineage Antibody Cocktail (Right Panel). Dead cells were excluded from analysis by staining with 7-AAD (7-Amino-Actinomycin D; Cat. No. 559925). Flow cytometry was performed on a BD LSR™II Flow Cytometry System.

Identification of CD34+ and CD34- subpopulations of hematopoietic progenitor cells. BALB/c bone marrow cells were treated with Mouse BD Fc Block™ Purified Rat anti-CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142) and stained with either FITC Rat IgG2a κ Isotype Control (Cat. No. 554688; data not shown) or FITC Rat anti-Mouse CD34 mAb RAM34 (Cat. No. 553733) and with either PerCP-Cy™ 5.5 Mouse Lineage Isotype Control Cocktail (Left Panel) or PerCP-Cy™ 5.5 Mouse Lineage Antibody Cocktail (Right Panel). Dead cells were excluded from analysis by staining with 7-AAD (7-Amino-Actinomycin D; Cat. No. 559925). Flow cytometry was performed on a BD LSR™II Flow Cytometry System.

Product Details
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BD Pharmingen™
Mouse (QC Testing)
Flow cytometry (Routinely Tested)
RUO
AB_10612020


Description

The PerCP-Cy™5.5 Mouse Lineage Antibody Cocktail has been designed to react with cells from the major hematopoietic lineages, such as T lymphocytes, B lymphocytes, monocytes/macrophages, NK cells, erythrocytes, and granulocytes. This pre-diluted Cocktail of five PerCP-Cy™5.5-conjugated antibodies is designed to label lineage marker-positive cells for exclusion to facilitate the flow cytometric identification of lineage marker-negative hematopoietic progenitors in mouse bone marrow. Components include clone 145-2C11, which recognizes Mouse CD3e; M1/70, which  recognizes CD11b; RA3-6B2, which recognizes CD45R/B220; TER-119, which recognizes Ly-76, mouse erythroid cells; and RB6-8C5, which recognizes Ly-6G and Ly-6C. PerCP-Cy™5.5 Mouse Lineage Isotype Control Cocktail contains equivalent concentrations of isotype-matched negative-control immunoglobulin. Additional fluorochrome-labeled reagents may be combined with the PerCP-Cy™5.5 Mouse Lineage Antibody Cocktail, and the PerCP-Cy™5.5 Mouse Lineage Isotype Control Cocktail, to further characterize hematopoietic progenitor subpopulations.

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

Recommended Assay Procedures

20 µl of the antibody or isotype control cocktail is sufficient to stain each sample of 10^6 leukocytes for flow cytometric analysis. We recommend the use of Mouse BD Fc Block™ purified anti-mouse CD16/Cd32 mAb 2.4G2 (Cat. No. 553141/553142) for optimal staining. The PerCP-Cy™5.5 Mouse Lineage Antibody Cocktail can be used to deplete cells bearing the hematopoietic lineage markers by flow cytometric sorting. The PerCP-Cy™5.5 fluorochrome is excited by laser lines from 595 to 647 nm, and its emission is collected in a detector for fluorescence wavelengths between 640 and 680 nm.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
  6. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  7. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  8. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
561317 Rev. 2
Components
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Description Quantity/Size Part Number EntrezGene ID
PerCP-Cy™5.5 Mouse Lineage Antibody Cocktail 100 Tests (1 ea) 51-9006964 N/A
PerCP-Cy™5.5 Mouse Lineage Isotype Control Cocktail 100 Tests (1 ea) 51-9006977 N/A
561317 Rev. 2
Citations & References
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Development References (6)

  1. Goodell MA, Rosenzweig M, Kim H, et al. Dye efflux studies suggest that hematopoietic stem cells expressing low or undetectable levels of CD34 antigen exist in multiple species. Nat Med. 1997; 3(12):1337-1345. (Biology). View Reference
  2. Morrison SJ, Wandycz AM, Hemmati HD, Wright DE, Weissman IL. Identification of a lineage of multipotent hematopoietic progenitors. Development. 1997; 124(10):1929-1939. (Biology). View Reference
  3. Okada S, Nakauchi H, Nagayoshi K, Nishikawa S, Miura Y, Suda T. In vivo and in vitro stem cell function of c-kit- and Sca-1-positive murine hematopoietic cells. Blood. 1992; 80(12):3044-3050. (Biology). View Reference
  4. 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.
  5. Spangrude GJ, Heimfeld S, Weissman IL. Purification and characterization of mouse hematopoietic stem cells. Science. 1988; 241(4861):58-62. (Biology). View Reference
  6. Spangrude GJ, Scollay R. A simplified method for enrichment of mouse hematopoietic stem cells. Exp Hematol. 1990; 18(8):920-926. (Biology). View Reference
View All (6) View Less
561317 Rev. 2

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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.