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Recombinant Mouse CD45 Protein, CF

Bio-Techne includes R&D Systems | Catalog # 114-CD

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114-CD-050

Key Product Details

Source

NS0

Accession #

Conjugate

Unconjugated

Applications

Binding Activity

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse CD45 protein
Gln24-Lys564 with a C-terminal 6-His tag

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

No results obtained: Gln24 predicted

Predicted Molecular Mass

59.5 kDa

SDS-PAGE

140-160 kDa, reducing conditions

Activity

Measured by its binding ability in a functional ELISA.
When 0.5 μg/mL of rmCD45 is captured onto a Mouse Anti-polyHistidine Monoclonal Antibody (Catalog # MAB050) coated plate, the concentration of rmGalectin-1 that produces 50% of the optimal binding is found to be approximately
0.45‑1.8 μg/mL.

Formulation, Preparation and Storage

114-CD
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution
Reconstitute at 100 μg/mL in PBS.

Reconstitution Buffer Available:
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Shipping The product is shipped at ambient temperature. Upon receipt, store it 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: CD45

CD45, previously called LCA (leukocyte common antigen), T200, or Ly5 in mouse, 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 mouse CD45RABC cDNA encodes 1291 amino acids (aa), including a 23 aa signal sequence, a 541 aa extracellular domain containing the splicing region, a cysteine-rich region and two fibronectin type III domains, a 22 aa transmembrane sequence, and a 705 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 30-169. 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

Alternate Names

CD45, LCA, PTPRC, T200 Glycoprotein

Entrez Gene IDs

5788 (Human); 19264 (Mouse); 490255 (Canine); 100061950 (Equine)

Gene Symbol

PTPRC

UniProt

Additional CD45 Products

Product Documents for Recombinant Mouse CD45 Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Mouse CD45 Protein, CF

For research use only

Customer Reviews for Recombinant Mouse CD45 Protein, CF (3)

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    Application: ELISA with mouse-specific anti CD45 proteins
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    Application: Stem/Immune cell maintenance or differentiation
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    Application: In vitro bioactivity in cell culture
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FAQs for Recombinant Mouse CD45 Protein, CF

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  • Q: This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?

    A: The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.

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