This monoclonal antibody recognizes a monomorphic determinant of the rat leukocyte common antigen.(1) The antigen recognized is a heavily glycosylated membrane glycoprotein of molecular weight 170,000 Da on thymocytes but molecular weight 170,000-220,000 Da on other leukocytes. The leukocyte common antigen (L-CA) is a major glycoprotein of haematopoietic cells but is not found on other tissues or erythroid cells. It is present on greater than 95 % of thymocytes, bone marrow cells and thoracic duct lymphocytes. This molecule carries much of the carbohydrate of thymocytes and shows interesting heterogeneity amongst T lymphocytes and B lymphocytes.
Flow Cytometry. Immunohistochemistry on Frozen Sections. (Reported to be unsuitable for use with Paraffin Sections). 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 with Lympholyte®-Rat 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. 2-1. 0 µg* of this Ab 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. 7. Wash 2 times at 4°C. 8. Add 100 µl of secondary antibody (Streptavidin-FITC) at a 1: 500 dilution. 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: Rat Strain: WistarCell Concentration: 1x10e6 cells per testAntibody Concentration Used: 0. 5 µg/10e6 cellsIsotypic Control: Biotin Mouse IgG2aCell Source Percentage of cells stained above control: Thymus: 99. 2%Spleen: 99. 2%Lymph Node: 99. 8%Results - Strain Distribution: Cell Concentration: 1x10e6 cells per testAntibody Concentration Used: 0. 5 µg/10e6 cellsStrains Tested: Wistar, Buffalo, Brown Norway, Fischer 344Positive: Wistar, Buffalo, Brown Norway, Fischer 344Negative none
限制
仅限研究用
浓度
0.1 mg/mL
缓冲液
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.
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.
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
CD45 (PTPRC)
(Protein tyrosine Phosphatase, Receptor Type, C (PTPRC))
CD45 is a family of single chain transmembraneous glycoproteins consisting of at least four isoforms (220, 205, 190, 180 kDa) which share a common large intracellular domain. Their extracellular domains are heavily glycosylated. The different isoforms are produced by alternative messenger RNA splicing of three exons of a single gene on chromosome 1. CD45 is expressed on cells of the human hematopoietic lineage (including hematopoietic stem cells) with the exception of mature red cells. It is not detected on differentiated cells of other tissues. It is likely that CD45 plays an important role in signal transduction, inhibition or upregulation of various immunological functions. Antibodies recognising a common epitope on all of the isoforms are termed CD45 whilst those recognising only individual isoforms are termed CD45RA or CD45RO etc.Synonyms: L-CA, Leukocyte common antigen, PTPRC, Receptor-type tyrosine-protein phosphatase C, T200