DESCRIPTION

PRAME (PReferentially expressed Antigen in MElanoma) is a tumor-associated antigen that was first identified through analysis of the specificity of tumor-reactive T-cell clones derived from a patient with metastatic cutaneous melanoma. It was subsequently found that PRAME is not only expressed in cutaneous melanoma, but also ocular melanoma and various nonmelanocytic malignant neoplasms, including non-small cell lung cancer, breast carcinoma, renal cell carcinoma, ovarian carcinoma, leukemia, synovial sarcoma, and myxoid liposarcoma. Normal healthy tissues are not known to express PRAME except for testis, ovary, placenta, adrenals, and endometrium. The expression of PRAME is a marker for an undifferentiated cellular state and has been implicated in the inhibition of tumour cell differentiation and the promotion of proliferation, which can contribute to oncogenesis.

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 8 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 16 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	8	94.99	7.51	7.91
2	8	22.47	1.67	7.44
3	8	6.25	0.50	7.97

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	89.91	9.66	10.74
2	16	21.87	1.81	8.27
3	16	6.21	0.50	8.08

RECOVERY

The recovery of human PRAME spiked to three different levels throughout the range of the assay in cell lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:32	72	71-74

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. (The cell lysate was initially diluted 1:2. )

		Cell lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	125
	Range (%)	123-127
1:8	Average% of Expected	124
	Range (%)	116-129
1:16	Average% of Expected	103
	Range (%)	81-116

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human PRAME Sandwich ELISA Kit 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
• Payment: T/T 100% in advance; PayPal; Alibaba Trade Assurance
• Shipping: FedEx / DHL / UPS express; cold-chain packing available on request
• Price: USD, per kit (48T / 96T options available)