Product Specifications for Recombinant Mouse CD45 Protein, CF
Mouse myeloma cell line, NS0-derived mouse CD45 protein Gln24-Lys564 with a C-terminal 6-His tag
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
<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
140-160 kDa, reducing conditions
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
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant
protein to be stored at a more dilute concentration.
The carrier free version does not contain BSA.
What formulation is right for me?
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or
as an ELISA standard.
In contrast, the carrier free protein is recommended for applications, in which the presence of BSA
Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitute at 100 μg/mL in PBS.
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.
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).
Anderson, J.N. et al. (2004) FASEB J. 18:8.
Streuli, M. et al. (1987) J. Exp. Med. 166:1548.
Hermiston, M.L. et al. (2003) Annu. Rev. Immunol. 21:107.
Earl, L.A. and L.G. Baum (2008) Immunol. Cell Biol. 86:608.
Ralph, S.J. et al. (1987) EMBO J. 6:1251.
Falahti, R. and D. Leitenberg (2008) J. Immunol. 181:6082.
Tchilian, E.Z. and P.C.L. Beverley (2006) Trends Immunol. 27:146.
McNiell, L. et al. (2007) Immunity 27:425.
Chen, I-J. et al. (2007) J. Biol. Chem. 282:35361.
Freiberg, B.A. et al. (2002) Nat. Immunol. 3:911.
Zhu, J.W. et al. (2008) Immunity 28:183.
Huntington, N.D. et al. (2006) Nat. Immunol. 7:190.
Hesslein, D.G. et al. (2006) Proc. Natl. Acad. Sci. USA 103:7012.
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?
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.
Product Documents for Recombinant Mouse CD45 Protein, CF
Which Brands are Currently Available on bio‑techne.com? R&D Systems, Tocris Bioscience and ProteinSimple branded products are available to purchase through bio‑techne.com. ProteinSimple branded instruments are available to quote. ACD branded products will be available on bio‑techne.com in the near future. Novus Biologicals branded products are not currently available on bio‑techne.com and can be found at novusbio.com.
Setting the standard in quality research reagents for over 30 years
A trusted leader in quality life science reagents
Your trusted supplier for innovative and high performance life science reagents
Proprietary systems and consumables for simpler, more quantitative and affordable protein analysis
Proprietary RNAscope® technology capable of detecting and quantifying RNA biomarkers in situ at single molecule sensitivity
A world leader in developing liquid biopsy based diagnostics