Key Product Details

Species Reactivity

Human

Applications

Immunohistochemistry, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 1066853
View all CD45RO Primary Antibodies »

Product Specifications

Immunogen

Chinese Hamster Ovary cell line, CHO-derived human CD45RO
Gln24-Lys391
Accession # P08575

Specificity

Detects rhCD45RO in Direct ELISA.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human CD45RO Antibody

Detection of CD45 in human PBMC cells (Positive) and Daudi cells (Negative).

CD45 was detected in immersion fixed human PBMC cells (Positive) and absent in Daudi human Burkitt's lymphoma cell line (Negative) using Mouse Anti-Human CD45RO Monoclonal Antibody (Catalog # MAB11594) 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 the cell membrane on a population of human PBMC's. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of CD45 in Human Tonsil.

CD45 was detected in immersion fixed paraffin-embedded sections of human tonsil using Mouse Anti-Human CD45RO Monoclonal Antibody (Catalog # mab11594) at 5 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 VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the cell surface. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Applications for Human CD45RO Antibody

Application
Recommended Usage

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed human PBMC cells and Daudi human Burkitt's lymphoma cell line

Immunohistochemistry

3-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CD45RO

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

CD45R, O isoform

Alternate Names

B220, CD45 antigen, CD45R, EC 3.1.3.48, L-CA, LY5, protein tyrosine phosphatase, receptor type, C, receptor-type tyrosine-protein phosphatase C, T200 glycoprotein, T200 leukocyte common antigen, T200receptor type, c polypeptide

Entrez Gene IDs

5788 (Human); 19264 (Mouse)

Gene Symbol

PTPRC

UniProt

P08575

Additional CD45RO Products

Product Documents for Human CD45RO Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human CD45RO Antibody

For research use only

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Protocols

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