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

Specifications

Product Data
Clone Name	RB6-8C5
Applications	FC, WB
Recommend Dilution	Flow cytometry (1,2,3). Western blot (5).
Reactivity	Mouse
Host	Rat
Clonality	Monoclonal
Specificity	This antibody reacts with the Mouse myeloid differentiation antigen GR-1 (1,2).
Formulation	PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml Label: FITC State: Liquid Ig fraction Absorption emission: 495 nm / 528 nm
Concentration	lot specific
Purification	Protein G chromatography
Conjugation	FITC
Storage Condition	Store the antibody at 2 - 8 °C for up to one month. For long term storage, aliquot and freeze unused portion at -20 °C in volumes appropriate for single usage. Avoid repeated freezing and thawing This product is photosensitive and should be protected from light.
Gene Name	lymphocyte antigen 6 complex, locus G
Database Link	Entrez Gene 546644 Mouse UniProt ID P35461
Background	GR-1 is a 25-30 kDa cell surface antigen and is expressed on myeloid cells but not lymphoid or erythroid 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 antibody labels: negative, low positive and high positive. Expression is transient on cells of monocytic lineage (3).
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. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add 0.1 - 0.5 µg of antibody per 10e6 cells. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. (It is recommended that the tubes are protected from light, since most fluorochromes are light sensitive.) 7. Wash 2 times at 4°C. 8. Resuspend the cell pellet in 50 µl ice cold media B. 9. 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: CBA/J Cell Concentration : 1x10e6 cells per test Antibody Concentration Used: 0.1 µg/10e6 cells Isotypic Control: FITC Rat IgG2b Cell Source Percentage of cells stained above control: Thymus 1.5% Whole Blood Monocytes 87.2% Bone Marrow Macrophages 90.0% (see picture below) Strain Distribution by Flow Cytometry Analysis: Procedure: see above Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 0.1 µg/10e6 cells Strains Tested: BALB/c, C57BL/6, CBA, C3H/he, AKR Positive: BALB/c, C57BL/6, CBA, C3H/he, AKR Negative: none
Reference Data

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