Human CD45 Antibody

Catalog # Availability Size / Price Qty
MAB14302-100
MAB14302-SP
CD45 in  Human PBMCs and MCF-7 Cell Line.
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Human CD45 Antibody Summary

Species Reactivity
Human
Specificity
Detects human CD45 in direct ELISA.
Source
Monoclonal Mouse IgG2A Clone # 998215
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human CD45
Thr24-Asn575
Accession # P08575
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
Immunohistochemistry
5-25 µg/mL
See below
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

Immunocytochemistry CD45 in  Human PBMCs and MCF-7 Cell Line. View Larger

CD45 in Human PBMCs and MCF-7 Cell Line. CD45 was detected in immersion fixed human peripheral blood mononuclear cells (positive staining; left panel) and MCF‑7 human breast cancer cell line (negative staining; right panel) using Mouse Anti-Human CD45 Monoclonal Antibody (Catalog # MAB14302) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Immunohistochemistry CD45  in Human Tonsil. View Larger

CD45 in Human Tonsil. CD45 was detected in immersion fixed paraffin-embedded sections of human tonsil using Mouse Anti-Human CD45 Monoclonal Antibody (Catalog # MAB14302) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

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Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Mouse IgG2A Isotype Control

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Reconstitution Buffers

Secondary Antibodies

Mouse IgG2A Antibody

Mouse IgG2A Antibody

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