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Ly-6G Neutrophil Marker antibody (FITC)

Details for Product No. ABIN114271, Supplier: Log in to see
Mouse (Murine)
Clonality (Clone)
Monoclonal ()
Flow Cytometry (FACS), Western Blotting (WB)
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Clone RB6-8C5
Isotype IgG2b
Characteristics Absorption / Emission: 495 nm / 528 nm
Purification Protein G chromatography
Alternative Name Ly6G / GR1 Neutrophil Marker
Background GR-1 is a 25-30 kDa cell surface antigen and is expressed on myeloid cells but not lymphoid or erythroid cells. The expression of the Gr-1 antigen increases with granulocyte maturation (3) as shown by the distinct populations of bone-marrow cells this monoclonal antibody labels: negative, low positive and high positive. Expression is transient on cells of monocytic lineage (3).Synonyms: Gr-1 Granulocyte marker
Gene ID 10090
UniProt P35461
Application Notes Flow cytometry (1,2,3). Western blot (5).
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add 0. 1 - 0. 5 µg of antibody per 10e6 cells. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. (It is recommended that the tubes areprotected from light, since most fluorochromes are light sensitive. )7. Wash 2 times at 4°C. 8. Resuspend the cell pellet in 50 µl ice cold media B. 9. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidiumiodide at 0. 5 mg/ml in PBS. This stains dead cells by intercalating in DNA. Media: A. Phosphate buffered saline (pH 7. 2) + 5% normal serum of host species + sodium azide(100 µl of 2M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7. 2) + 0. 5% Bovine serum albumin + sodium azide (100µl of 2M sodium azide in 100 mls). Results: Tissue Distribution by Flow Cytometry Analysis: Mouse Strain: CBA/JCell Concentration : 1x10e6 cells per testAntibody Concentration Used: 0. 1 µg/10e6 cellsIsotypic Control: FITC Rat IgG2bCell Source Percentage of cells stained above control: Thymus 1. 5%Whole Blood Monocytes 87. 2%Bone Marrow Macrophages 90. 0%(see picture below)Strain Distribution by Flow Cytometry Analysis: Procedure: see aboveCell Concentration: 1x10e6 cells per test
Restrictions For Research Use only
Concentration 0.1 mg/mL
Buffer PBS, 0.02 % NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/mL
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Avoid repeated freezing and thawing. Dilute only prior to immediate use
Storage 4 °C/-20 °C
Storage Comment Store the antibody (undiluted) at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
Supplier Images
Flow Cytometry (FACS) image for anti-Ly-6G Neutrophil Marker antibody (FITC) (ABIN114271) Rat anti Granulocytes (Gr-1 antigen) RB6-8C5
Background publications Jutila, Kroese, Jutila, Stall, Fiering, Herzenberg, Berg, Butcher: "Ly-6C is a monocyte/macrophage and endothelial cell differentiation antigen regulated by interferon-gamma." in: European journal of immunology, Vol. 18, Issue 11, pp. 1819-26, 1989 (PubMed).

Spangrude, Heimfeld, Weissman: "Purification and characterization of mouse hematopoietic stem cells." in: Science (New York, N.Y.), Vol. 241, Issue 4861, pp. 58-62, 1988 (PubMed).

Brummer, Sugar, Stevens: "Immunological activation of polymorphonuclear neutrophils for fungal killing: studies with murine cells and blastomyces dermatitidis in vitro." in: Journal of leukocyte biology, Vol. 36, Issue 4, pp. 505-20, 1984 (PubMed).