Ly-6G Neutrophil Marker antibody (FITC)

Antigen: Ly-6G Neutrophil Marker

Clonality: Monoclonal (RB6-8C5)

Host: Rat

Reactivity: Mouse (Murine)

Conjugate: FITC

Application: Western Blotting (WB), Flow Cytometry (FACS)

Catalog no.: ABIN114271

Additional Information

Alternative name: Ly6G / GR1 Neutrophil Marker

UniProt: P35461

Cross-Reactivity: Mouse (Murine)

Isotype: IgG2b

Clone: RB6-8C5

Description: GR-1 is a 25-30 kDa cell surface antigen and is expressed on myeloid cells but not lymphoidor 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 antibodylabels: negative, low positive and high positive. Expression is transient on cells ofmonocytic lineage (3).

Characteristics: Synonyms: Gr-1 Granulocyte marker

Application Details

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

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.

Concentration: 0.1 mg/ml

Buffer: PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring totalprotein concentration to 4-5 mg/mlLabel: FITC Absorption / Emission: 495 nm / 528 nm

Storage: Store the antibody at 2 - 8 °C for up to one month. For long term storage, aliquot and freezeunused portion at -20 °C in volumes appropriate for single usage. Avoid repeated freezingand thawing This product is photosensitive and should be protected from light. Shelf life: one year from despatch.

Restrictions: For Research Use only

Images

Rat anti Granulocytes (Gr-1 antigen) RB6-8C5

Publications

Jutila, Kroese, Jutila et al.: "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).