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Recombinant Human SIRP alpha / CD172a Protein (HPLC-verified), His tag, 1mg  

Recombinant Human SIRP alpha / CD172a Protein (HPLC-verified), His tag, 1mg

Recombinant Human SIRP alpha / CD172a Protein (Glu 31 - Arg 370), produced in human 293 cells (HEK293), HPLC-verified, His tag

Synonym
Recombinant Human Protein, SHPS1, SIRPA, CD172A, BIT, MFR, MYD1, P84, PTPNS1

More details

SIA-H5225-1K

Availability: within 7 days

1 980,00 €

Background
Tyrosine-protein phosphatase non-receptor type substrate 1 (SHPS1) 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.

Source
Recombinant Human SIRP alpha / CD172a Protein (HPLC-verified) (Human SIRP alpha (HPLC-verified), His Tag) Glu 31 - Arg 370 (Accession # NP_001035111) was produced in human 293 cells (HEK293).

Molecular Characterization
Human SIRP alpha (HPLC-verified), His Tag is fused with a polyhistidine tag at the C-terminus, and has a calculated MW of 38.1 kDa. The predicted N-terminus is Glu 31. The reducing (R) protein migrates as 45-55 kDa in SDS-PAGE due to glycosylation.

Endotoxin
Less than 1.0 EU per μg of the Human SIRP alpha (HPLC-verified), His Tag by the LAL method.

Purity
>95% as determined by SDS-PAGE.
>95% as determined by SEC-HPLC.

Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally Mannitol or Trehalose are added as protectants before lyophilization.

Reconstitution
See Certificate of Analysis for details of reconstitution instruction and specific concentration.

Storage
Avoid repeated freeze-thaw cycles. No activity loss was observed after storage at:
In lyophilized state for 1 year (4°C-8°C); After reconstitution under sterile conditions for 1 month (4°C-8°C) or 3 months (-20°C to -70°C).

Bioactivity-ELISA
Serial dilutions of Anti-Human CD47 Neutralizing Antibody were added into Human SIRP alpha, His Tag (HPLC-verified) (Cat. No. SIA-H5225): Biotinylated Human CD47, Fc,Avitag (Cat. No. CD7-H82F6) binding reactions. The half maximal inhibitory concentration (IC50) is 0.1245 μg/mL (Routinely tested).

Bioactivity-BLI
Loaded Human CD47, Mouse IgG2a Fc Tag, low endotoxin (Cat. No. CD7-H52A5) on Protein A Biosensor, can bind Human SIRP alpha, His Tag (HPLC-verified) (Cat. No. SIA-H5225) with an affinity constant of 1.1 μM as determined in BLI assay (ForteBio Octet Red96e) (QC tested).
Immobilized Human CD47, Fc Tag (HPLC-verified) (Cat. No. CD7-H5256) on AHC Biosensor, can bind Human SIRP alpha, His Tag (HPLC-verified) (Cat. No. SIA-H5225) with an affinity constant of 0.72 μM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Bioactivity-FACS
FACS assay shows that Human SIRP alpha, His Tag (HPLC-verified) (Cat. No. SIA-H5225) can bind to Jurkat cell expressing CD47. The concentration of SIRP alpha used is 1 μg/mL (Routinely tested).
FACS analysis shows that the binding of Human SIRP alpha, His Tag (HPLC-verified) (Cat. No. SIA-H5225) to Jurkat expressing CD47 was inhibited by increasing concentration of neutralizing Anti-CD47 antibody. The concentration of SIRP alpha used is 1 μg/mL. IC50=0.2257 μg/mL (Routinely tested).

References

(1) Timms J.F., et al., 1999, Curr. Biol. 9:927-930.
(2) Stofega M.R., et al., 2000, J. Biol. Chem. 275:28222-28229.
(3) Latour S., et al., 2001, J. Immunol. 167:2547-2554.