DESCRIPTION

FLNA encodes a 280 kDa widely expressed actin binding protein called filamin A which can crosslinks actin filaments and links actin filaments to membrane glycoproteins to form a three-dimensional network. And filamin A could interact with many other proteins, involved in receptor activation, cell migration and adhesion, cell proliferation, inflammation and tumorigenesis, studies have been reported that Filamin A overexpressed in multiple types of tumors, suggesting Filamin A may involved in cancer aggressiveness. What's more, Filamin A is an causative gene of periventricular nodular heterotopia (PNH).

PRINCIPLE OF THE ASSAY

An anti-tag antibody is pre-coated onto the bottom of wells. After adding antigen or samples, Tagged-Capture antibody and HRP-conjugated detection antibody, a sandwich complex is formed in the solution. TMB acts as a HRP substrate, and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns the 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	2,610.4	55.8	2.1
2	8	628.2	10.8	1.7
3	8	326.8	8.2	2.5

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	2,614.8	46.7	1.8
2	16	631.6	9.3	1.5
3	16	334.0	19.6	5.9

RECOVERY

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

Sample Type		Average% of Expected	Range (%)
Human serum	1:200	108	94-122
	1:400	104	102-105
Cell lysate	1:50	88	85-90
	1:100	93	88-101

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 human serum was initially diluted 1:25. The cell lysate was initially diluted 1:100.)

		Human serum	Cell lysate
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	89	96
	Range (%)	87-90	82-109
1:8	Average% of Expected	93	97
	Range (%)	86-100	80-114
1:16	Average% of Expected	88	100
	Range (%)	83-93	81-119

CALIBRATION

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