DESCRIPTION

CD16, also known as the low affinity Fc gamma receptor III for IgG (FcγRIII), exists in two isoforms, FcγRIIIa (CD16a) and FcγRIIIb (CD16b), encoded by two nearly identical genes, FCGR3A and the FCGR3B. CD16a is expressed on NK cells, macrophages, and placental trophoblasts as a polypeptide-anchored transmembrane protein while CD16b is expressed on neutrophils in a glycosylphosphatidylinositol (GPI)-anchored form. CD16a forms a heteromeric structure with the Fc epsilon RI (gamma) and/or CD3 (zeta) subunits. CD16 mediates antibody-dependent cellular cytotoxicity (ADCC) and other antibody-dependent responses, such as phagocytosis. CD16 can be cleaved and released as a soluble form (sCD16).

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with HRP) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with HRP also binds to the analyte. TMB acts as the HRP substrate and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns solution yellow. The color intensity is proportional to the quantity of bound protein which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 20 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 24 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	20	12.58	0.43	3.5
2	20	3.07	0.08	2.7
3	20	0.82	0.04	5.0

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	24	13.24	0.49	3.7
2	24	3.22	0.11	3.5
3	24	0.79	0.06	7.0

RECOVERY

The recovery of human CD16 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human serum	1:80	107	94-124
	1:160	84	71-105

LINEARITY

To assess the linearity of the assay, human serum samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. (The human serum samples were initially diluted 1:20 )

		Human serum
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	107
	Range (%)	97-125
1:8	Average% of Expected	103
	Range (%)	96-112
1:16	Average% of Expected	92
	Range (%)	86-104

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human CD16 produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
• Lead time: varies with order quantity and specifications. Please consult your local distributor or our sales team for confirmation
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• Shipping: FedEx / DHL / UPS express; cold-chain packing available on request
• Price: USD, per kit (48T / 96T options available)