Flow cytometry (1,2,3). Western blot (5). Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
实验流程
FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation. 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. 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 areprotected 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 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 by Flow Cytometry Analysis: Mouse Strain: CBA/JCell Concentration : 1x10e6 cells per testAntibody Concentration Used: 0. 1 µg/10e6 cellsIsotypic Control: FITC Rat IgG2bCell 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 aboveCell Concentration: 1x10e6 cells per test
限制
仅限研究用
浓度
0.1 mg/mL
缓冲液
PBS, 0.02 % NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/mL
储存液
Sodium azide
注意事项
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
注意事项
Avoid repeated freezing and thawing. Dilute only prior to immediate use
储存条件
4 °C/-20 °C
储存方法
Store the antibody (undiluted) at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
抗原
Ly6g
(Lymphocyte Antigen 6 Complex, Locus G (Ly6g))
Gr-1 antibody, Gr1 antibody, Ly-6G antibody, lymphocyte antigen 6 complex, locus G antibody, Ly6g antibody
背景
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