CD45 / LCA (CD45RB) Rat Monoclonal Antibody [Clone ID: 16A]

CAT#: CL029P

CD45 / LCA (CD45RB) rat monoclonal antibody, clone 16A, Purified

Product Images

Cell source: Spleen. Percentage of cells stained above control: 63.0%

Specifications

Product Data

• Clone Name: 16A
• Applications: FC, IHC, IP
• Recommended Dilution: Immunohistochemistry on acetone-fixed frozen sections and paraffin embedded sections.
  Immunoprecipitation.
  Flow cytometry (for details see "Specificity" and "Protocols" below).
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: TH2 cell clones, final boost with TH2 clone D10
• Specificity: Anti-CD45RB monoclonal antibody reacts with the CD45 isoform containing the exon B dependent epitope.
  CD45RB is highly expressed on peripheral B cells, cytotoxic T cells, a subset of T helper cells and most thymoctyes.
  Tissue distribution by flow cytometry analysis:
  Mouse strain: BALB/c
  Cell concentration : 1x10e6 cells per test
  Antibody dilution used: 50 µl at 1/5000/10e6 cells
  Isotypic control: Rat IgG2a
  Cell source / Percentage of cells stained above control:
  Thymus / 81.1%
  Spleen / 63.0%
  Lymph Node / 74.1%
• Formulation: PBS containing 0.02% sodium azide
  State: Purified
  State: Liquid purified Ig fraction
• Concentration: 1.0 mg/ml
• Purification: Protein G chromatography
• Background: 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: PTPRC, Leukocyte common antigen, L-CA, T200
• 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 1x10e6 cells, representing 1 test).
  4. To each tube, add 50 µl of a 1/2500-1/5000 dilution of CL029P.
  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-rat IgG (H+L)) at a ~ 1/500 dilution.
  9. Incubate the tubes at 4°C for 30-60 minutes.
  (It is recommended that the tubes are protected 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 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).