CD45 antibody

Catalog No. ABIN114375

Product Details anti-CD45 Antibody

Target: CD45 (PTPRC) (Protein tyrosine Phosphatase, Receptor Type, C (PTPRC))

Reactivity: Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: This CD45 antibody is un-conjugated

Application: Flow Cytometry (FACS), Immunohistochemistry (Frozen Sections) (IHC (fro))

Specificity: This monoclonal antibody recognizes a monomorphic determinant of the rat leukocyte common antigen.

Purification: Protein G Chromatography

Immunogen: Lymph node glycoproteins and cells. Donor: BALB/c spleen Fusion Partner: NSO/U

Clone: OX-30

Isotype: IgG2a

Application Details

Application Notes: Flow cytometry (see protocol). Immunohistochemistry with frozen sections.
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 Lympholyte®-Rat cell separation medium. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50µl of thissuspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add 0. 5-1. 0 µg* of this Ab. 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-mouse IgG (H+L)) at 1: 700 dilution. 9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes areprotected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C in media B. 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: Mouse IgG2aCell Source Percentage of cells stained above control: Thymus: 99. 9%Spleen: 97. 4%Lymph Node: 90. 6%Results - Strain Distribution: Antibody Concentration Used: 0. 5 µg/10e6 cellsStrains Tested: Wistar, Buffalo, Brown Norway, Fischer 344

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

Buffer: PBS and 0.02 % NaN3

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.

Target Details for CD45

Target: CD45 (PTPRC) (Protein tyrosine Phosphatase, Receptor Type, C (PTPRC))

Alternative Name: CD45 / LCA (PTPRC Products)

Synonyms: PTPRC antibody, si:dkeyp-47c5.1 antibody, B220 antibody, CD45 antibody, CD45R antibody, GP180 antibody, L-CA antibody, LCA antibody, LY5 antibody, T200 antibody, Cd45 antibody, Ly-5 antibody, Lyt-4 antibody, loc antibody, Lca antibody, RT7 antibody, cd45 antibody, protein tyrosine phosphatase, receptor type C antibody, protein tyrosine phosphatase, receptor type, C antibody, receptor-type tyrosine-protein phosphatase C antibody, PTPRC antibody, ptprc antibody, LOC100074271 antibody, Ptprc antibody

Background: The antigen recognized is a heavily glycosylated membrane glycoprotein of molecular weight 170,000 kDa on thymocytes but molecular weight 170,000-220,000 kDa 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 lymphocytesSynonyms: L-CA, Leukocyte common antigen, PTPRC, Receptor-type tyrosine-protein phosphatase C, T200

Gene ID: 24699

NCBI Accession: NP_001103357

UniProt: P04157

Pathways: TCR Signaling, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, CXCR4-mediated Signaling Events, BCR Signaling