CD45RO Antibody (1066853) [Unconjugated]

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.

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.

• Reactivity: Hu
• Applications: IHC, ICC/IF
• Clone: 1066853
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

CD45RO Antibody (1066853) [Unconjugated] Summary

• Immunogen: Chinese Hamster Ovary cell line, CHO-derived human CD45RO
  Gln24-Lys391
  Accession # P08575
• Specificity: Detects rhCD45RO in Direct ELISA.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 3-25 ug/mL
  • Immunohistochemistry 3-25 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Reconstitution Instructions: Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CD45RO Antibody (1066853) [Unconjugated]

• B220
• CD45 antigen
• CD45R
• CD45RO
• 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

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

FAQs for CD45RO Antibody (MAB11594). (Showing 1 - 1 of 1 FAQ).

1. I cannot find any anti-mouse CD45RO for FACS. Have any of your anti-Human CD45RO been tested on mouse T cells?
   • Unfortunately, none of our CD45RO antibodies for Flow Cytometry have been validated on mouse. If you would be interested in testing the antibody on mouse, we would recommend Our Innovators Reward Program. Under the terms and conditions of this program we would be happy to provide you with a credit of equal or lesser value, in exchange for your online review including an image of either positive or negative staining for testing in a new species and/or application. Please contact us at innovators@novusbio.com with any questions regarding this program.

Bioinformatics

• Uniprot:
  • P08575()