Cell-based Assay Verified Siglec-15 Protein

Siglec-15 is a member of the sialic acid-binding Ig-like lectin (Siglecs) family and is expressed in M2 macrophages, myeloid cells, dendritic cells, B cells, and osteoclasts. Siglec-15 can continuously inhibit the activity of T cells and exhibits the main features of normalized cancer immunotherapy. Related studies have shown that Siglec-15 is the major immunosuppressive factor in most PD-L1 negative tumor patients. And the anti-cancer effect of this target is likely to be mutually exclusive with PD-L1. In tumor cells, the expression level of siglec-15 is low when PD-L1 expression is high. For those cancer patients who are not effective in PD-1/PD-L1 treatment, Siglec-15 may bring them a new choice of immunotherapy.

High bioactivity Siglec-15 protein can help drugs development. The bioactivity of Siglec-15 protein developed by ACROBiosystems is verified by Cell based assay/BLI /SPR /ELISA, and the corresponding protocols are provided for free.

The bioactivity of Siglec-15 proteins are verified by Cell based assay/ BLI/ SPR/ ELISA, and the protocols are offered.

Cell based assay

Biotinylated Human Siglec-15, Fc,Avitag (Cat. No. SG5-H82F5) inhibits Anti-CD3-induced proliferation of PBMC. The ED50 for this effect is 0.48-0.57 μg/mL.

Protocol

BLI

Loaded Anti-Siglec-15 MAb (Mouse IgG) on AMC Biosensor, can bind Human Siglec-15, Fc Tag (Cat. No. SG5-H5253) with an affinity constant of 59.7 pM as determined in BLI assay (ForteBio Octet Red96e).

Protocol

SPR

Immobilized Neu5Ac(a2-6)GalNAc-PAA-biotin on SA Chip can bind Human Siglec-15, Fc Tag (Cat. No. SG5-H5253) with an affinity constant of 4.49 μM as determined in a SPR assay (Biacore T200).

Protocol

ELISA

Immobilized Mouse Human chimeric MAb (5G12, Human IgG1) at 5 μg/mL (100 μL/well) can bind Biotinylated Human Siglec-15, Fc,Avitag (Cat. No. SG5-H82F5) with a linear range of 0.4-3 ng/mL.

Protocol

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