DESCRIPTION

Rad50, an ATPase, binds to Mre11 and Nbs1 to form a repair complex that is transported to the nucleus in response to DNA double-strand breaks. Rad50 is a component of MRN (Mre11-Rad50-Nbs1), which participates in DNA double-strand break repair and DNA-damage checkpoint activation. Rad50 promoted proliferation/invasion and induced EMT of ovarian cancer cells, whereas knockdown of Rad50 led to decreased aggressive behaviors. Frequent lack of RAD50 was found in endometrial and low-grade epithelial ovarian cancer, whereas Rad50 was highly expressed in melanoma and gastric cancer.

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 (pg/mL)	SD	CV%
1	8	2,573.2	69.6	2.7
2	8	615.1	42.8	7.0
3	8	327.8	9.1	2.8

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	2,606.1	91.6	3.5
2	16	612.6	34.3	5.6
3	16	324.5	16.5	5.1

RECOVERY

The recovery of human RAD50 spiked to three different levels throughout the range of the assay in cell lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:8	93	78-104

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	97
	Range (%)	93-100
1:8	Average% of Expected	95
	Range (%)	92-98
1:16	Average% of Expected	82
	Range (%)	81-83

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human RAD50 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
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• Price: USD, per kit (48T / 96T options available)