Human SIRP alpha /CD172a Antibody AF4546: R&D Systems

Human SIRP alpha /CD172a Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human SIRP alpha /CD172a in Western blots.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Chinese hamster ovary cell line CHO-derived recombinant human SIRP alpha /CD172a
    Gly27-Asn370
    Accession # P78324
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    50 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human SIRP alpha /CD172a by Western Blot. Western blot shows lysates of U2OS human osteosarcoma cell line, A549 human lung carcinoma cell line, and U937 human histiocytic lymphoma cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human SIRP alpha /CD172a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4546) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for SIRP alpha /CD172a at approximately 52 kDa (unglycosylated) and 90‑120 kDa (glycosylated) as indicated. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human SIRP alpha /CD172a by Simple WesternTM. Simple Western lane view shows lysates of U2OS human osteosarcoma cell line and A549 human lung carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for SIRP alpha /CD172a at approximately 113-116 kDa (as indicated) using 50 µg/mL of Sheep Anti-Human SIRP alpha /CD172a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4546) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
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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