Ly-6G Neutrophil Marker antibody (Biotin)

Antigen: Ly-6G Neutrophil Marker

Clonality: Monoclonal (RB6-8C5)

Host: Rat

Reactivity: Mouse (Murine)

Conjugate: Biotin

Application: Flow Cytometry (FACS)

Catalog no.: ABIN114270

Additional Information

Alternative name: Ly6G / GR1 Neutrophil Marker

UniProt: P35461

Immunogen: Mouse Granulocytes.

Cross-Reactivity: Mouse (Murine)

Isotype: IgG2b

Clone: RB6-8C5

Characteristics: Synonyms: Gr-1 Granulocyte marker

Specificity: Anti-mouse Gr-1 monoclonal antibody reacts with the myeloid differentiation antigen Gr-1. (1,2). This 25-30 kDa cell surface antigen is expressed on myeloid cells but not lymphoid orerythroid 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). ABIN114273 is a useful antibody for studies of myeloid differentiation stages and theirregulations by cytokines.

Application Details

Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with cell separation medium. 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. 2 µg* of ABIN114273. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. 7. Wash 2 times at 4°C. 8. Add 100 µl of secondary antibody FITC Goat anti-rat IgG (H+L) . 9. Incubate tubes at 4°C for 30-60 minutes (It is recommended that tubes are protectedfrom light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C. 11. Resuspend the cell pellet in 50 µl ice cold media B. 12. 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: Mouse Strain: BALB/cCell

Application Notes: Flow cytometry. Immunohistochemistry on frozen and paraffin embedded tissues. Western blot. Complement-mediated depletion. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.

Concentration: 1x10e6 cells per testAntibody Concentration Used: 0.1 µg/10e6 cellsIsotypic Control: Rat IgG2bCell: Source Percentage of cells stained above control: Thymus: 3.5%Whole Blood: Granulocytes 96.1%Monocytes: 97.7%Bone Marrow: Granulocytes 92.1%Monocytes: 94.1

Buffer: PBS, 0.02% sodium azide as preservative and EIA grade BSA as astabilizing protein to bring total protein concentration to 4-5 mg/ml.Label: Biotin

Storage: Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing. Shelf life: one year from despatch.

Restrictions: For Research Use only

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).

Whiteland, Nicholls, Shimeld et al.: "Immunohistochemical detection of T-cell subsets and other leukocytes in paraffin-embedded rat and mouse tissues with monoclonal antibodies." in: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, Vol. 43, Issue 3, pp. 313-20, 1995 (PubMed).