分子别名(Synonym)
CD47,MER6,IAP,OA3
表达区间及表达系统(Source)
Human CD47, Fc Tag (CD7-H5256) is expressed from human 293 cells (HEK293). It contains AA Gln 19 - Pro 139 (Accession # Q08722-3).
Predicted N-terminus: Gln 19
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蛋白结构(Molecular Characterization)
This protein carries a human IgG1 Fc tag at the C-terminus.
The protein has a calculated MW of 40.4 kDa. The protein migrates as 55-66 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
内毒素(Endotoxin)
Less than 1.0 EU per μg by the LAL method.
纯度(Purity)
>95% as determined by SDS-PAGE.
>90% as determined by SEC-MALS.
制剂(Formulation)
Lyophilized from 0.22 μm filtered solution in 50 mM Tris, 100 mM Glycine, pH7.5 with trehalose as protectant.
Contact us for customized product form or formulation.
重构方法(Reconstitution)
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
存储(Storage)
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 3 months under sterile conditions after reconstitution.
电泳(SDS-PAGE)
Human CD47, Fc Tag on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95%.
SEC-MALS
The purity of Human CD47, Fc Tag (Cat. No. CD7-H5256) is more than 90% and the molecular weight of this protein is around 95-110 kDa verified by SEC-MALS.
Report
活性(Bioactivity)-ELISA
Immobilized Human CD47, Fc Tag (Cat. No. CD7-H5256) at 2 μg/mL (100 μL/well) can bind Human SIRP alpha, His Tag (Cat. No. SIA-H5225) with a linear range of 0.016-0.5 μg/mL (QC tested).
Protocol
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).
Protocol
活性(Bioactivity)-BLI
Immobilized Human CD47, Fc Tag (Cat. No. CD7-H5256) on AHC Biosensor, can bind Human SIRP alpha, His Tag (Cat. No. SIA-H5225) with an affinity constant of 0.72 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
Protocol
活性(Bioactivity)-FACS
FACS assay shows that Human CD47, Fc Tag (Cat. No. CD7-H5256) can bind to ACHN cell expressing SIRP-a. The concentration of CD47 used is 10 μg/mL (Routinely tested).
Protocol
FACS analysis shows that the binding of Human CD47 to ACHN expressing SIRP-a was inhibited by increasing concentration of neutralizing SIRP-a antibody. The concentration of Human CD47 used is 10 μg/mL. IC50=15.02 μg/mL (Routinely tested).
Protocol
背景(Background)
Leukocyte surface antigen CD47 is also known as Antigenic surface determinant protein OA3, Integrin-associated protein (IAP) and Protein MER6. CD47 contains 1 Ig-like V-type (immunoglobulin-like) domain. CD47 is very broadly distributed on normal adult tissues. CD47 has a role in both cell adhesion by acting as an adhesion receptor for THBS1 on platelets, and in the modulation of integrins and plays an important role in memory formation and synaptic plasticity in the hippocampus by similarity. CD47 is the receptor for SIRPA, binding to which prevents maturation of immature dendritic cells and inhibits cytokine production by mature dendritic cells. CD47 Interaction with SIRPG mediates cell-cell adhesion, enhances superantigen-dependent T-cell-mediated proliferation and costimulates T-cell activation.