Human CD45 Antibody

(5 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human CD45. This antibody recognizes all isoforms of human CD45.
  • Source
    Monoclonal Mouse IgG1 Clone # 2D1
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Human peripheral blood mononuclear cells
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • Immunohistochemistry
    5-25 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    8-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of CD45 in Human Blood Lymphocytes by Flow Cytometry. Human peripheral blood lymphocytes were stained with Mouse Anti-Human CD45 Monoclonal Antibody (Catalog # MAB1430, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). View our protocol for Staining Membrane-associated Proteins.
Immunocytochemistry
CD45 in Human PBMCs. CD45 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with PHA blast using Mouse Anti-Human CD45 Monoclonal Antibody (Catalog # MAB1430) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; Catalog # NL007) and counter­stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Immunohistochemistry
CD45 in Hodgkin's Lymphoma Tissue. CD45 was detected in immersion fixed paraffin-embedded sections of Hodgkins lymphoma tissue using Mouse Anti-Human CD45 Monoclonal Antibody (Catalog # MAB1430) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surfaces. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: CD45

CD45, previously called LCA (leukocyte common antigen), T200, or Ly5 in mice, is member C of the class 1 (receptor‑like) protein tyrosine phosphatase family (PTPRC) (1, 2). It is a variably glycosylated 180‑220 kDa transmembrane protein that is abundantly expressed on all nucleated cells of hematopoietic origin (1‑3). CD45 has several isoforms, expressed according to cell type, developmental stage and antigenic exposure (1‑5). The longest form, CD45RABC (called B220 in mouse), is expressed on B lymphocytes (5). The CD45RABC cDNA encodes 1304 amino acids (aa), including a 23 aa signal sequence, a 552 aa extracellular domain containing the splicing region, a cysteine‑rich region and two fibronectin type III domains, a 22 aa transmembrane sequence, and a 707 aa cytoplasmic domain that contains two phosphatase domains, D1 and D2. Only D1 has phosphatase activity. CD45R0 is the shortest form, lacking exons 4, 5 and 6 which encode aa 32‑191. It is expressed on memory cells, while intermediate sizes are expressed on other T cells (3, 4, 6). CD45 has been best studied in T cells, where it determines T cell receptor signaling thresholds (3, 6‑8). CD45 is moved into or out of the immunological synapse (IS) membrane microdomain depending on the relative influence of interaction with the extracellular galectin lattice or the intracellular actin cytoskeleton (9, 10). Galectin interaction can be fine‑tuned by varying usage of the heavily O‑glycosylated spliced regions and sialylation of N‑linked carbohydrates (4, 9). Within the IS, CD45 dephosphorylates and negatively regulates the Src family kinase, Lck (8‑10). In other leukocytes, CD45 influences differentiation and links immunoreceptor signaling with cytokine secretion and cell survival, partially overlapping in function with DEP‑1/CD148 (11‑14). CD45 deletion causes in severe immunodeficiency, while point mutations may be associated with autoimmune disorders (6, 7).

  • References:
    1. Anderson, J.N. et al. (2004) FASEB J. 18:8.
    2. Streuli, M. et al. (1987) J. Exp. Med. 166:1548.
    3. Hermiston, M.L. et al. (2003) Annu. Rev. Immunol. 21:107.
    4. Earl, L.A. and L.G. Baum (2008) Immunol. Cell Biol. 86:608.
    5. Ralph, S.J. et al. (1987) EMBO J. 6:1251.
    6. Falahti, R. and D. Leitenberg (2008) J. Immunol. 181:6082.
    7. Tchilian, E.Z. and P.C.L. Beverley (2006) Trends Immunol. 27:146.
    8. McNiell, L. et al. (2007) Immunity 27:425.
    9. Chen, I-J. et al. (2007) J. Biol. Chem. 282:35361.
    10. Freiberg, B.A. et al. (2002) Nat. Immunol. 3:911.
    11. Zhu, J.W. et al. (2008) Immunity 28:183.
    12. Huntington, N.D. et al. (2006) Nat. Immunol. 7:190.
    13. Hesslein, D.G. et al. (2006) Proc. Natl. Acad. Sci. USA 103:7012.
    14. Cross, J.L. et al. (2008) J. Immunol. 180:8020.
  • Long Name:
    Cluster of Differentiation 45
  • Entrez Gene IDs:
    5788 (Human); 19264 (Mouse); 490255 (Canine); 100061950 (Equine)
  • Alternate Names:
    B220; CD45 antigen; CD45; CD45R; EC 3.1.3.48; LCA; L-CA; LY5; protein tyrosine phosphatase, receptor type, C; PTPRC; receptor-type tyrosine-protein phosphatase C; T200 Glycoprotein; T200 leukocyte common antigen; T200receptor type, c polypeptide
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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Species
Applications
Sample Type
  1. Preferential Lineage-Specific Differentiation of Osteoblast-Derived Induced Pluripotent Stem Cells into Osteoprogenitors
    Authors: CL Roberts, SS Chen, AC Murchison, RA Ogle, MP Francis, RC Ogle, PC Sachs
    Stem Cells Int, 2017;2017(0):1513281.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow
  2. Proteomic profiling of peripheral blood neutrophils identifies two inflammatory phenotypes in stable COPD patients
    Authors: ALT Loi, S Hoonhorst, C van Aalst, J Langereis, V Kamp, S Sluis-Eisi, N Ten Hacken, JW Lammers, L Koenderman
    Respir. Res., 2017;18(1):100.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow
  3. Feasibility and Efficiency of Human Bone Marrow Stromal Cell Culture with Allogeneic Platelet Lysate-Supplementation for Cell Therapy against Stroke
    Stem Cells Int, 2016;2016(0):6104780.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow
  4. Switch from canonical to noncanonical Wnt signaling mediates high glucose-induced adipogenesis.
    Authors: Keats E, Dominguez J, Grant M, Khan Z
    Stem Cells, 2014;32(6):1649-60.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
  5. Isolation and characterization of synovial mesenchymal stem cells.
    Authors: Harvanova D, Tothova T, Sarissky M, Amrichova J, Rosocha J
    Folia Biol. (Praha), 2011;57(3):119-24.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
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