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
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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).
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