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Ly-6G Neutrophil Marker antibody
Alternatives Cytotoxicity Test (CyTox), Western Blotting (WB), Flow Cytometry (FACS)
|5 references available|
|Quantity||0.5 mL (1x10e6 cells per testAntibody Conce...) (Variants)|
|Price||528.00 $ Plus shipping costs $45.00|
|Availability||Will be delivered in 6 to 8 Business Days|
|Alternative name||Ly6G / GR1 Neutrophil Marker|
|Characteristics||Synonyms: Gr-1 Granulocyte marker|
|Protocol||FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with Lympholyte®-M 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 50 µl of a 1: 5000-1: 10000 dilution* of this AB. 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)) at a 1: 500 dilution. 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 - Tissue Distribution: Mouse Strain: BALB/cCell|
|Application Notes||Flow Cytometry. Complement mediated depletion. Western blot. Immunohistochemistry on Frozen and Paraffin Sections. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.|
|Concentration||1x10e6 cells per testAntibody Concentration Used: 50µl of a 1: 5000 dilution/10e6 cellsIsotypic Control: Rat IgG2bCell Source Percentage of cells stained above control: Thymus: 9.7%Whole Blood - Granulocytes: 98.6%Whole Blood - Monocytes: 93.5%Bone Marrow|
|Storage||Prior to reconstitution and Following reconstitution store the antibody undiluted at 2-8°Cfor 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|
|Cell Source: Bone Marrow Monocytes - Percentage of cells stained above control: 93.2%|
Hestdal, Ruscetti, Ihle et al.: "Characterization and regulation of RB6-8C5 antigen expression on murine bone marrow cells." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 147, Issue 1, pp. 22-8, 1991 (PubMed).
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).
|Hosts||Rat (14), Mouse (1)|
|Reactivities||Mouse (Murine) (15)|
|Applications||Flow Cytometry (FACS) (11), Immunohistochemistry (Frozen Sections) (IHC (fro)) (7), Functional Studies (Func) (6), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) (6), Western Blotting (WB) (6), ELISA (5), Immunoprecipitation (IP) (2), Cytotoxicity Test (CyTox) (1)|
|Conjugates||PE (4), Biotin (3), FITC (3)|