Ly6g Rat Monoclonal Antibody [Clone ID: RB6-8C5]

Specifications

Product Data
Clone Name	RB6-8C5
Applications	FC, IHC, WB
Recommend Dilution	Flow Cytometry (Ref.1-3). Immunohistochemistry on Frozen and Paraffin Embedded Sections (Ref.6). Western blot (Ref.5). Complement-mediated depletion.
Reactivity	Mouse
Host	Rat
Clonality	Monoclonal
Immunogen	Mouse Granulocytes.
Specificity	Anti-Mouse Gr-1 monoclonal antibody reacts with the myeloid differentiation antigen Gr-1. (Ref.1,2).  This 25-30 kDa cell surface antigen is expressed on myeloid cells but not lymphoid or erythroid cells. The expression of the Gr-1 antigen increases with granulocyte maturation (Ref.3) as shown by the distinct populations of bone-marrow cells this monoclonal antibody labels: negative, low positive and high positive. Expression is transient on cells of monocytic lineage (Ref.3).
Formulation	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 Label: Biotin State: Liquid Ig fraction
Concentration	lot specific
Purification	Protein G Chromatography
Conjugation	Biotin
Storage Condition	Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing.
Gene Name	lymphocyte antigen 6 complex, locus G
Database Link	Entrez Gene 546644 Mouse UniProt ID P35461
Synonyms	Gr-1 Granulocyte marker
Note	Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with cell separation medium. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x107 cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1 x 106 cells, representing 1 test). 4. To each tube, add 0.1-0.2 µg* of CL046B per 106 cells. 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 Streptavidin at 1/500 dilution. 9. Incubate tubes at 4°C for 30-60 minutes (It is recommended that tubes are protected from 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 propidium iodide 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 (100 µl of 2M sodium azide in 100 mls). Results: Tissue Distribution by Flow Cytometry Analysis: Mouse Strain: BALB/c Cell Concentration: 1x106 cells per test Antibody Concentration Used: 0.1 µg/106 cells Isotypic Control: Rat IgG2b Cell: Source Percentage of cells stained above control: Thymus: 1.5% Whole Blood: Granulocytes: 90.8%, Monocytes: 91.8% Bone Marrow: Granulocytes: 94.2%, Monocytes: 95.3% Cell Source: Bone Marrow Monocytes Percentage of cells stained above control: 95.3% Strain Distribution by Flow Cytometry Analysis: Cell Concentration: 1x106 cells per test. Antibody Concentration Used: 0.1 µg/106 cells. Strains Tested: BALB/c, C57BL/6, CBA, C3H/He. Positive: BALB/c, C57BL/6, CBA, C3H/He. Negative: none.
Reference Data

*Delivery time may vary from web posted schedule. Occasional delays may occur due to unforeseen complexities in the preparation of your product. International customers may expect an additional 1-2 weeks in shipping.