Biotinylated Human SIRP alpha, Fc,Avitag (CDA-H82F2) is expressed from human 293 cells (HEK293). It contains AA Glu 31 - Arg 370 (Accession # P78324-1).
Predicted N-terminus: Glu 31
This protein carries a human IgG1 Fc tag at the C-terminus, followed by a Avi tag (Avitag™).
The protein has a calculated MW of 65.8 kDa. The protein migrates as 75-105 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
The biotin to protein ratio is 0.5-1 as determined by the HABA assay.
Less than 1.0 EU per μg by the LAL method.
>95% as determined by SDS-PAGE.
>90% as determined by SEC-MALS.
Lyophilized from 0.22 μm filtered solution in Tris with Glycine, Arginine and NaCl, pH7.5. Normally trehalose is added as protectant before lyophilization.
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Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 12 months under sterile conditions after reconstitution.
Biotinylated Human SIRP alpha, Fc,Avitag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
The purity of Biotinylated Human SIRP alpha, Fc,Avitag(Cat. No. CDA-H82F2) was more than 90% and the molecular weight of this protein is around 155-170 kDa verified by SEC-MALS.
Immobilized Human CD47, His Tag (Cat. No. CD7-H5227) at 5 μg/mL (100 μL/well) can bind Biotinylated Human SIRP alpha, Fc,Avitag (Cat. No. CDA-H82F2) with a linear range of 5-205 ng/mL (QC tested).
Serial dilutions of Anti-Human CD47 Neutralizing Antibody were added into Human CD47, Fc Tag (Cat. No. CD7-H5256): Biotinylated Human SIRP alpha, Fc,Avitag (Cat. No. CDA-H82F2) binding reactions. The half maximal inhibitory concentration (IC50) is 0.5431 μg/mL (Routinely tested).
FACS assay shows that Biotinylated Human SIRP alpha, Fc,Avitag (Cat. No. CDA-H82F2) can bind to Jurkat cell expressing CD47. The concentration of SIRP alpha used is 3 μg/mL (QC tested).
FACS analysis shows that the binding of Biotinylated Human SIRP alpha, Fc,Avitag (Cat. No. CDA-H82F2) to Jurkat expressing CD47 was inhibited by increasing concentration of neutralizing Anti- Human CD47 antibody. The concentration of SIRP alpha used is 1 μg/mL. IC50=0.1303 μg/mL (Routinely tested).
Tyrosine-protein phosphatase non-receptor type substrate 1 (SHPS1) is also known as CD172 antigen-like family member A (CD172a), Macrophage fusion receptor, MyD-1 antigen, Signal-regulatory protein alpha (SIRPA or SIRP alpha) or p84, is a member of the SIRP family, and also belongs to the immunoglobulin superfamily. SIRP alpha is Ubiquitous and highly expressed in brain. SIRPA / CD172a is immunoglobulin-like cell surface receptor for CD47 and acts as docking protein and induces translocation of PTPN6, PTPN11 and other binding partners from the cytosol to the plasma membrane. SIRPA / SHPS-1 supports adhesion of cerebellar neurons, neurite outgrowth and glial cell attachment and may play a key role in intracellular signaling during synaptogenesis and in synaptic function By similarity. SIRPA / MyD1 involved in the negative regulation of receptor tyrosine kinase-coupled cellular responses induced by cell adhesion, growth factors or insulin and mediates negative regulation of phagocytosis, mast cell activation and dendritic cell activation. CD47 binding prevents maturation of immature dendritic cells and inhibits cytokine production by mature dendritic cells.