Recombinant Human SIRP alpha Fc Biotinylated Protein, CF

    
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  • Purity
    >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
  • Endotoxin Level
    <0.10 EU per 1 μg of the protein by the LAL method.
  • Activity
    Measured by the ability of the immobilized protein to support the adhesion of human red blood cells. Subramanian, S. et al. (2006) Blood 107:2548. Red blood cells, added at 10 x 106 cells/mL (100 µL/well) to SIRP alpha coated plates, will adhere after 1 hour at 37 °C. The ED50 for this effect is 0.6-2.4 µg/mL.
    Optimal dilutions should be determined by each laboratory for each application.
  • Source
    Chinese Hamster Ovary cell line, CHO-derived human SIRP alpha/CD172a protein
    Human SIRP alpha
    (Gly27-Arg370) &
    (Glu31-Arg370)
    Accession # NP_542970
    IEGRMDHuman IgG1
    (Pro100-Lys330)
    N-terminus
  • Accession #
  • N-terminal Sequence
    Analysis
    Gly27 & Glu31
  • Structure / Form
    Disulfide-linked homodimer. Biotinylated via sugars
  • Predicted Molecular Mass
    64 kDa (unlabeled)
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BT4546
 
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
 
Reconstitution Reconstitute at 500 μg/mL in PBS.
 
Shipping The product is shipped with polar packs. 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.
 
Data Image
Both immobilized unlabeled Recombinant Human SIRP alpha /CD172a Fc Chimera (Catalog # 4546-SA) and Biotinylated Recombinant Human SIRP alpha/CD172a Fc Chimera (Catalog # BT4546) support the adhesion of human red blood cells. The ED50 for this effect is 0.6-2.4 μg/mL. The similarity in activity highlights that the biotinylated protein is fully functional.
Background: SIRP alpha/CD172a
Signal regulatory protein alpha (SIRP alpha, designated CD172a), also called SHPS-1 (SHP substrate 1) and previously, MyD-1 (Myeloid/Dendritic-1), is a monomeric ~90 kDa type I transmembrane glycoprotein that belongs to the SIRP/SHPS (CD172) family of the immunoglobulin superfamily (1-4). SIRPs are paired receptors, with similar extracellular domains but differing C-termini and functions (1, 2). The 503 amino acid (aa) human SIRP alpha contains a 342 aa extracellular domain (ECD), with one V-type, and two C1 type Ig domains, and three potential N glycosylation sites. It has a 110 aa cytoplasmic sequence with ITIM motifs that recruit tyrosine phosphatases SHP-1 and SHP-2 when phosphorylated (4). Human SIRP alpha has more than 40 described polymorphisms, including the prominent BIT (Brain Ig like molecule with Tyrosine-based activation motifs, also called SIRP alpha 2 or PTPNS) (5). One reported isoform lacks aa 1-101, which eliminates most of the V type Ig domain. Human SIRP alpha ECD shares 61%, 60%, 71%, 72% and 73% aa identity with mouse, rat, porcine, bovine and equine SIRP alpha, respectively; it shares 84% and 76% aa identity with human SIRP beta 1 and SIRP gamma, respectively (2). SIRP alpha is expressed mainly on myeloid cells, including macrophages, neutrophils, dendritic and Langerhans cells (3-6). It is also found on neurons, smooth muscle and endothelial cells (7-9). SIRP alpha shows adhesion to the ubiquitous CD47/IAP (integrin associated protein), while SIRP gamma binds more weakly and SIRP alpha 1 does not bind at all (1, 2). Mouse and human SIRP alpha -CD47 binding only cross-reacts for specific polymorphisms and influences engraftment of xenotransplanted stem cells (6, 10). SIRP alpha engagement generally produces a negative regulatory signal (4). Low SIRP alpha recognition of CD47, which occurs on aged erythrocytes or platelets or xenogenic cells, promotes clearance of CD47low cells from circulation (11, 13). SIRP alpha recognition of surfactants SP-A and SP-D in the lung can inhibit alveolar macrophage cytokine production (14). The CD47 integrin-SIRP alpha interaction is reported to promote macrophage fusion during osteoclastogenesis (15).
  • References:
    1. Barclay, A.N. & M.H. Brown (2006) Nat. Rev. Immunol. 6:457.
    2. vanBeek, E.M. et al. (2005) J. Immunol. 175:7781.
    3. Liu, Y. et al. (2005) J. Biol. Chem. 280:36132.
    4. Kharitonenkov, A. et al. (1997) Nature 386:181.
    5. Swissprot Accession # P78324.
    6. Miyashita, M. et al. (2004) Mol. Biol. Cell 15:3950.
    7. Wang, X.X. & K.H. Pfenninger (2005) J. Cell Sci. 119:172.
    8. Maile, L.A. et al. (2003) Mol. Biol. Cell 14:3519.
    9. Johansen, M.L. & E.J. Brown (2007) J. Biol. Chem. 282:24219.
    10. Takenaka, K. et al. (2007) Nat. Immunol. 8:1313.
    11. Ishikawa-Sekigami, T. et al. (2006) Biochem. Biophys. Res. Commun. 343:1197.
    12. Olsson, M. et al. (2005) Blood 105:3577.
    13. Ide, K. et al. (2007) Proc. Natl. Acad. Sci. USA 104:5062.
    14. Gardai, S.J. et al. (2003) Cell 115:13.
    15. Lundberg, P. et al. (2007) Biochem. Biophys. Res. Commun. 352:444.
  • Long Name:
    Signal-regulatory Protein alpha
  • Entrez Gene IDs:
    140885 (Human); 19261 (Mouse); 25528 (Rat); 101926317 (Cynomolgus Monkey)
  • Alternate Names:
    BIT; BITbrain-immunoglobulin-like molecule with tyrosine-based activation motifs; Brain Ig-like molecule with tyrosine-based activation motifs; CD172 antigen-like family member A; CD172a antigen; CD172a; Inhibitory receptor SHPS-1; Macrophage fusion receptor; MFR; MFRtyrosine phosphatase SHP substrate 1; MyD-1 antigen; MYD1; MYD-1; P84; protein tyrosine phosphatase, non-receptor type substrate 1; PTPNS1; SHP substrate 1; SHPS1; SHPS-1; SHPS1CD172A; signal-regulatory protein alpha; Signal-regulatory protein alpha-1; Signal-regulatory protein alpha-2; Signal-regulatory protein alpha-3; SIRP alpha; SIRPA; SIRPalpha; Sirp-alpha-1; SIRPalpha2; Sirp-alpha-2; Sirp-alpha-3; SIRPtyrosine-protein phosphatase non-receptor type substrate 1
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