Recombinant Human SIRP alpha/CD172a Fc Avi-tag Protein, CF

Catalog #: AVI4546 Datasheet / COA / SDS

Biotinylated

Catalog #	Availability	Size / Price	Qty
AVI4546-050

Recombinant Human SIRP alpha/CD172a Fc Avi-tag Protein Binding Activity

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All SIRP alpha/CD172a Proteins and Enzymes

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human SIRP alpha/CD172a Fc Avi-tag Protein, CF Summary

Learn more about Avi-tag Biotinylated Proteins

Product Specifications

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

The biotin to protein ratio is greater than 0.7 as determined by the HABA assay. Measured by its binding ability in a functional ELISA. When Recombinant Human CD47 Fc Chimera (Catalog # 4670-CD) is immobilized at 0.2 µg/mL (100 µL/well), Biotinylated Recombinant Human SIRP alpha /CD172a Fc Chimera Avi-tag (Catalog # AVI4546) binds with an ED₅₀ of 20-180 ng/mL.

Source

Chinese Hamster Ovary cell line, CHO-derived human SIRP alpha/CD172a protein

Human SIRP alpha /CD172a
(Glu31-Asn371)
Accession # P78324.2

IEGRMD

Human IgG₁
(Pro100-Lys330)

Avi-tag

Accession #

P78324.2

N-terminal Sequence
Analysis

Glu31

Structure / Form

Disulfide-linked homodimer, biotinylated via Avi-tag

Predicted Molecular Mass

66 kDa

SDS-PAGE

86-96 kDa, under reducing conditions

Product Datasheets

AVI4546

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. 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 could interfere.

AVI4546

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 500 μg/mL in PBS.
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.

Scientific Data

Binding Activity

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When Recombinant Human CD47 Fc Chimera (Catalog # 4670-CD) is immobilized at 0.2 µg/mL (100 µL/well), Biotinylated Recombinant Human SIRPa/CD172a Fc Chimera Avi-tag (Catalog # AVI4546) binds with an ED₅₀ of 20-180 ng/mL.

SDS-PAGE

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2 μg/lane of Biotinylated Recombinant Human SIRP alpha /CD172a Fc Chimera Avi-tag (Catalog # AVI4546) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 86-96 kDa and 170- 195 kDa, respectively.

Reconstitution Calculator

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 CD47^lowcells 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

Barclay, A.N. & M.H. Brown (2006) Nat. Rev. Immunol. 6:457.
vanBeek, E.M. et al. (2005) J. Immunol. 175:7781.
Liu, Y. et al. (2005) J. Biol. Chem. 280:36132.
Kharitonenkov, A. et al. (1997) Nature 386:181.
Swissprot Accession # P78324.
Miyashita, M. et al. (2004) Mol. Biol. Cell 15:3950.
Wang, X.X. & K.H. Pfenninger (2005) J. Cell Sci. 119:172.
Maile, L.A. et al. (2003) Mol. Biol. Cell 14:3519.
Johansen, M.L. & E.J. Brown (2007) J. Biol. Chem. 282:24219.
Takenaka, K. et al. (2007) Nat. Immunol. 8:1313.
Ishikawa-Sekigami, T. et al. (2006) Biochem. Biophys. Res. Commun. 343:1197.
Olsson, M. et al. (2005) Blood 105:3577.
Ide, K. et al. (2007) Proc. Natl. Acad. Sci. USA 104:5062.
Gardai, S.J. et al. (2003) Cell 115:13.
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); 494566 (Porcine); 609452 (Canine); 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