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Ly-6G/Ly-6C Monoclonal Antibody (RB6-8C5), NovaFluor™ Red 700, eBioscience™
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FIGURE: 1 / 3
Ly-6G/Ly-6C Antibody (M016T02R03) in Flow
C57BL/6 mouse splenocytes were unstained (left) or stained with 0.05 µg of Ly-6G/Ly-6C Monoclonal Antibody, NovaFluor Red 700 (Product # m016t02r03) (right). All cells were co-stained with CD45R (B220) (RA3-6B2) Monoclonal Antibody, eFluor 450 (Product # 48-0452-82). Total viable cells were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora and unmixed with autofluorescence extraction.
Product Details
M016T02R03
Product Specifications
Host/Isotype
Rat
/ IgG2b, kappa
Class
Monoclonal
Type
Antibody
Clone
RB6-8C5
Conjugate
Excitation/Emission Max
637/701 nm
View spectra
Form
Liquid
Concentration
0.1 mg/mL
Amount
25 µg
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
Product Specific Information
Description
The RB6-8C5 monoclonal antibody reacts with mouse Ly-6G, a 21-25 kDa protein also known as the myeloid differentiation antigen Gr-1. A GPI-linked protein, Gr-1 is expressed by the myeloid lineage in a developmentally regulated manner in the bone marrow. While monocytes only express Gr-1 transiently during their bone marrow development, the expression of Gr-1 on bone marrow granulocytes as well as on peripheral neutrophils is a good marker for these populations. eBioscience testing indicates that in the bone marrow and lysed whole blood, the antibody clone RB6-8C5 also stains cells that express the highest levels of Ly6c (as defined by staining with antibody clone HK1.4). It is recommended that 1A8-Ly6G (Product # 9668) be used when looking at Ly-6G specific targets.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
The RB6-8C5 antibody has been tested by flow cytometric analysis of mouse bone marrow cells and splenocytes. This can be used at less than or equal to 0.05 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 µL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 µL of CellBlox Plus Blocking Buffer per 100 µL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Excitation: 637 nm; Emission: 701 nm; Laser: 633-640 nm (Red) Laser
Target Information
Granulocytes are a category of white blood cells characterised by the presence of granules in their cytoplasm. They are also called polymorphonuclear leukocytes (PMN or PML) because of the varying shapes of the nucleus, which is usually lobed into three segments. In common parlance, the term polymorphonuclear leukocyte often refers specifically to neutrophil granulocytes, the most abundant of the granulocytes. Granulocytes or PMN are released from the bone marrow by the regulatory complement proteins.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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