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Ly-6G Neutrophil Marker antibody (Biotin)

Details for Product No. ABIN114269, Supplier: Log in to see
Antigen
Reactivity
Mouse (Murine)
13
Host
Rat
13
Clonality (Clone)
Monoclonal ()
Conjugate
Biotin
3
3
2
Application
Flow Cytometry (FACS), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
10
5
5
5
4
4
2
1
Supplier
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Immunogen Mouse Granulocytes.
Clone RB6-8C5
Isotype IgG2b
Cross-Reactivity (Details) Species reactivity (tested):Mouse.
Alternative Name Ly6G / GR1 Neutrophil Marker
Background Synonyms: Gr-1 Granulocyte marker
Gene ID 10090
UniProt P35461
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.
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
Restrictions For Research Use only
Buffer PBS containing 0.02 % Sodium Azide as preservative 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.
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.
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).