Ly-6G Neutrophil Marker antibody

Details for Product No. ABIN114273, Supplier: Log in to see
Mouse (Murine)
Clonality (Clone)
Monoclonal ()
Cytotoxicity Test (CyTox), Flow Cytometry (FACS), Western Blotting (WB)
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Clone RB6-8C5
Isotype IgG2b
Purification Protein G Chromatography.
Alternative Name Ly6G / GR1 Neutrophil Marker
Background Synonyms: Gr-1 Granulocyte marker
Gene ID 10090
UniProt P35461
Application Notes This antibody is suitable for studies of myeloid differentiation stages and their regulationsby cytokines. Applications include Flow Cytometry (1,2,3) complement-mediated depletion(4), Western blot staining (5) and both Frozen and Paraffin Sections (6).
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 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-Tissue Distribution: Mouse Strain: BALB/cCell Concentration: 1x10e6 cells per testAntibody Concentration Used: 0. 1 µg/10e6 cellsIsotypic Control: Rat IgG2bCell Source-Percentage of cells stained above control: Thymus: 2. 0%Whole Blood Monocytes: 79. 1%Bone Marrow Macrophages: 87. 5%N. B. Appropriate control samples should always be included in any labelling studies. * For optimal results in various applications, it is recommended that each investigatordetermine dilutions appropriate for individual use. Strain Distribution by Flow Cytometry Analysis: Cell Concentration: 1x10e6 cells per testAntibody Concentration Used: 0. 1 µg/10e6 cellsStrains Tested: BALB/c, C57BL/6, CBA, C3H/He, AKR
Restrictions For Research Use only
Concentration 1.0 mg/mL
Buffer PBS and 0.02 % Sodium Azide as preservative.
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.
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 (ABIN114273) Rat anti Granulocytes (Gr-1 antigen) RB6-8C5
Background publications 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).

Muller-Sieburg, Whitlock, Weissman: "Isolation of two early B lymphocyte progenitors from mouse marrow: a committed pre-pre-B cell and a clonogenic Thy-1-lo hematopoietic stem cell." in: Cell, Vol. 44, Issue 4, pp. 653-62, 1986 (PubMed).